Derivatives of 5-(hetero)arylpyrazol-3-carboxylic amide or 1-(hetero)aryltriazol-4-carboxylic amide useful for the treatment of inter alia cystic fibrosis

ABSTRACT

The present disclosure is based, in part, on the discovery that disclosed compounds such as those having Formula (IVa), (Va), (IV), or (V) can increase cystic fibrosis transmembrane conductance regulator (CFTR) activity as measured in human bronchial epithelial (hBE) cells.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a national stage filing under 35 U.S.C. § 371 ofPCT/US2015/000211, filed Dec. 23, 2015, which claims the benefit of, andpriority to, U.S. provisional application Ser. No. 62/102,227, filedJan. 12, 2015, and 62/096,384, filed Dec. 23, 2014, the contents of eachof which are hereby incorporated by reference herein in their entirety.

BACKGROUND

Cells normally maintain a balance between protein synthesis, folding,trafficking, aggregation, and degradation, referred to as proteinhomeostasis, utilizing sensors and networks of pathways (Sitia et al.,Nature 426: 891-894, 2003; Ron et al., Nat Rev Mol Cell Biol 8: 519-529,2007). The cellular maintenance of protein homeostasis, or proteostasis,refers to controlling the conformation, binding interactions, locationand concentration of individual proteins making up the proteome. Proteinfolding in vivo is accomplished through interactions between the foldingpolypeptide chain and macromolecular cellular components, includingmultiple classes of chaperones and folding enzymes, which minimizeaggregation (Wiseman et al., Cell 131: 809-821, 2007). Whether a givenprotein folds in a certain cell type depends on the distribution,concentration, and subcellular localization of chaperones, foldingenzymes, metabolites and the like (Wiseman et al.). Cystic fibrosis andother maladies of protein misfolding arise as a result of an imbalancein the capacity of the protein homeostasis (proteostasis) environment tohandle the reduced energetic stability of misfolded, mutated proteinsthat are critical for normal physiology (Balch et al., Science 319,916-9 (2008); Powers, et al., Annu Rev Biochem 78, 959-91 (2009); Huttet al., FEBS Lett 583, 2639-46 (2009)).

Cystic Fibrosis (CF) is caused by mutations in the cystic fibrosistransmembrane conductance regulator (CFTR) gene which encodes amulti-membrane spanning epithelial chloride channel (Riordan et al.,Annu Rev Biochem 77, 701-26 (2008)). Approximately ninety percent ofpatients have a deletion of phenylalanine (Phe) 508(ΔF508) on at leastone allele. This mutation results in disruption of the energetics of theprotein fold leading to degradation of CFTR in the endoplasmic reticulum(ER). The ΔF508mutation is thus associated with defective folding andtrafficking, as well as enhanced degradation of the mutant CFTR protein(Qu et al., J Biol Chem 272, 15739-44 (1997)). The loss of a functionalCFTR channel at the plasma membrane disrupts ionic homeostasis (Cl⁻,Na⁺, HCO₃ ⁻) and airway surface hydration leading to reduced lungfunction (Riordan et al.). Reduced periciliary liquid volume andincreased mucus viscosity impede mucociliary clearance resulting inchronic infection and inflammation, phenotypic hallmarks of CF disease(Boucher, J Intern Med 261, 5-16 (2007)). In addition to respiratorydysfunction, ΔF508CFTR also impacts the normal function of additionalorgans (pancreas, intestine, gall bladder), suggesting that theloss-of-function impacts multiple downstream pathways that will requirecorrection.

In addition to cystic fibrosis, mutations in the CFTR gene and/or theactivity of the CFTR channel has also been implicated in otherconditions, including for example, congenital bilateral absence of vasdeferens (CBAVD), acute, recurrent, or chronic pancreatitis,disseminated bronchiectasis, asthma, allergic pulmonary aspergillosis,smoking-related lung diseases, such as chronic obstructive pulmonarydisease (COPD), dry eye disease, Sjogren's syndrome and chronicsinusitis, (Sloane et al. (2012), PLoS ONE 7(6):e39809.doi:10.1371/journal. pone.0039809; Bombieri et al. (2011), J CystFibros. 2011 Jun. 10Suppl 2:S86-102; (Albert et al. (2008). ClinicalRespiratory Medicine, Third Ed., Mosby Inc.; Levin et al. (2005), InvestOphthalmol Vis Sci., 46(4):1428-34; Froussard (2007), Pancreas 35(1):94-5).

There remains a need in the art for compounds, compositions and methodsof increasing CFTR activity as well as for methods of treating CF, otherCFTR-related diseases, and other maladies of protein misfolding.

SUMMARY

The present disclosure is based, in part, on the discovery thatdisclosed compounds can increase cystic fibrosis transmembraneconductance regulator (CFTR) activity as measured in human bronchialepithelial (hBE) cells.

For example, provided herein, in an embodiment, are compounds having theFormula (IV) or (V):

and pharmaceutically acceptable salts, stereoisomers, and prodrugsthereof, wherein:

X₁ is CH or N; wherein when X₁ is N, R^(z) is not present and thenitrogen with R^(z) has a double bond with the adjacent N and there is asingle bond between X₁ and the adjacent N, and when X₁ is CH, X₁ has adouble bond with the nitrogen adjacent, and IV is selected from thegroup consisting of H, and C₁₋₃alkyl, and the nitrogen with IV has asingle bond with the adjacent N;

X₃ is selected from the group consisting of O, S, and NR_(hh);

pp is 1, 2, or 3;

R₁₁ is independently selected for each occurrence from the groupconsisting of hydrogen, halogen, C₁₋₄alkyl (optionally substituted byone, two or three halogens);

L₁ is selected from the group consisting of C₁₋₆ alkylene,C₃₋₆cycloalkylene, C₃₋₆cycloalkylene-C₁₋₄alkylene,C₁₋₃alkylene-NR_(hh)-S(O)_(w-), —C₁₋₃ alkylene-S(O)_(w)—NR_(hh)—,C₃₋₆cycloalkylene-C₀₋₂ alkylene-S(O)_(w)—NR_(hh), and C₃₋₆cycloalkylene-C₀₋₂alkylene NR_(hh)-S(O)_(w-), wherein L₁ may be optionally substitutedby one, two or three substituents selected from the group consisting ofhalogen, hydroxyl, C₁₋₃alkyl (optionally substituted by one, two orthree substituents each selected independently from R_(ff));

R₄₄ is selected from the group consisting of H, halogen, hydroxyl,C₁₋₃alkoxy, heterocycle, and a 5-6 membered monocyclic or 8-10memberedbicyclic heteroaryl having one, two or three heteroatoms each selectedfrom O, N, and S; wherein the heterocycle and the heteroaryl may beoptionally substituted by one or two substituents each selectedindependently from R_(gg);

R_(ff) is selected for each occurrence from group consisting of halogen,hydroxyl, C₁₋₄alkyl, C₁₋₄alkyoxy, C₂₋₄alkenyl, C₃₋₆cycloalkyl, —NR′R″,—NR′—S(O)_(w)—C₁₋₃alkyl, S(O)_(w)—NR′R″, and —S(O)_(w)—C₁₋₃alkyl, wherew is 0, 1, or 2, wherein C₁₋₄alkyl, C₁₋₄alkyoxy, C₂₋₄alkenyl andC₃₋₆cycloalkyl may be optionally substituted by one, two or threesubstituents each independently selected from the group consisting ofhalogen, hydroxyl, —NR′R″, —NR′—S(O)_(w)—C₁₋₃alkyl, S(O)_(w)—NR′R″, and—S(O)_(w)—C₁₋₃alkyl;

R_(gg) is selected for each occurrence from group consisting of halogen,hydroxyl, C₁₋₆alkyl, C₁₋₆alkyoxy, C₂₋₆alkenyl, C₃₋₆cycloalkyl, —NR′R″,—NR′—S(O)_(w)—C₁₋₃alkyl, S(O)_(w)—NR′R″, and —S(O)_(w)—C₁₋₃alkyl, wherew is 0, 1, or 2, wherein C₁₋₆alkyl, C₁₋₆alkyoxy, C₂₋₆alkenyl andC₃₋₆cycloalkyl may each be optionally substituted by one, two or threesubstituents each independently selected from the group consisting ofhalogen, C₁₋₆alkyl, C₁₋₆alkoxy, hydroxyl, C(O)OH, —C(O)OC₁₋₆alkyl,—O—C₃₋₆cycloalkyl, —O-heterocycle, —O-heteroaryl, —O-phenyl, —NR′R″,—NR′—S(O)_(w)—C₁₋₃alkyl, S(O)_(w)—NR′R″, and —S(O)_(w)—C₁₋₃alkyl;

w is 0, 1 or 2; and

R_(hh) is selected for each occurrence from the group consisting of H,C₁₋₆alkyl and C₃₋₆cycloalkyl.

Also contemplated herein are pharmaceutical compositions that include adisclosed compound such as those compounds having Formula (IVa), (Va),(IV), or (V) and a pharmaceutically acceptable carrier or excipient. Incertain embodiments, the compositions can include at least oneadditional CFTR modulator as described anywhere herein or at least twoadditional CFTR modulators, each independently as described anywhereherein.

In additional embodiments, a method of enhancing (e.g., increasing)cystic fibrosis transmembrane conductance regulator (CFTR) activity in asubject in need thereof is provided comprising administering to saidsubject an effective amount of a compound of Formula (IVa), (Va), (IV),or (V).

In certain of these embodiments, the activity of one or more (e.g., oneor two) mutant CFTRs (e.g., ΔF508, S549N, G542X, G551D, R117H, N1303K,W1282X, R553X, 621+1G>T, 1717−1G>A, 3849+10kbC>T, 2789+5G>A, 3120+1G>A,I507del, R1162X, 1898+1G>A, 3659delC, G85E, D1152H, R560T, R347P,2184insA, A455E, R334W, Q493X, and 2184delA CFTR) is enhanced (e.g.,increased). In certain embodiments, ΔF508CFTR activity is enhanced(e.g., increased). In other embodiments, the activities of two mutantCFTRs (e.g., ΔF508and G551D; ΔF508and A455E; or G542X; Δ508F) areenhanced (e.g., increased).

In certain of these embodiments, the subject (e.g., a human patient) issuffering from a disease associated with decreased CFTR activity (e.g.,cystic fibrosis, congenital bilateral absence of vas deferens (CBAVD),acute, recurrent, or chronic pancreatitis, disseminated bronchiectasis,asthma, allergic pulmonary aspergillosis, chronic obstructive pulmonarydisease (COPD), chronic sinusitis, dry eye disease, protein Cdeficiency, A-β-lipoproteinemia, lysosomal storage disease, type 1chylomicronemia, mild pulmonary disease, lipid processing deficiencies,type 1 hereditary angioedema, coagulation-fibrinolyis, hereditaryhemochromatosis, CFTR-related metabolic syndrome, chronic bronchitis,constipation, pancreatic insufficiency, hereditary emphysema, Sjogren'ssyndrome, familial hypercholesterolemia, I-cell disease/pseudo-Hurler,mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II,polyendocrinopathy/hyperinsulemia, Diabetes mellitus, Laron dwarfism,myleoperoxidase deficiency, primary hypoparathyroidism, melanoma,glycanosis CDG type 1, congenital hyperthyroidism, osteogenesisimperfecta, hereditary hypofibrinogenemia, ACT deficiency, Diabetesinsipidus (DI), neurophyseal DI, nephrogenic DI, Charcot-Marie Toothsyndrome, Perlizaeus-Merzbacher disease, Alzheimer's disease,Parkinson's disease, amyotrophic lateral sclerosis, progressivesupranuclear palsy, Pick's disease, Huntington's disease,spinocerebellar ataxia type I, spinal and bulbar muscular atrophy,dentatorubral pallidoluysian, myotonic dystrophy, hereditaryCreutzfeldt-Jakob disease (due to prion protein processing defect),Fabry disease, and Straussler-Scheinker syndrome). In certainembodiments, the disease is cystic fibrosis.

In yet additional aspects, the disclosure is directed to treating apatient suffering from cystic fibrosis comprising administering to saidpatient an effective amount of a disclosed compound.

In some embodiments, the methods described herein can further includeadministering an additional CFTR modulator or administering at least twoadditional CFTR modulators. In certain embodiments, at least one CFTRmodulator is a CFTR corrector (e.g., VX-809, VX-661, VX-983, VX-152,VX-440, GLPG2222 and GLPG2665) or potentiator (e.g., ivacaftor andgenistein). In certain of these embodiments, one of the at least twoadditional therapeutic agents is a CFTR corrector (e.g., VX-809, VX-661,VX-983, VX-152, VX-440, GLPG2222 and GLPG2665) and the other is a CFTRpotentiator (e.g., ivacaftor and genistein).

In a further aspect, a method of identifying a candidate agent thatincreases CFTR activity is provided, which includes: (i) contacting acell that expresses a CFTR protein with the candidate agent and adisclosed compound; (ii) measuring the CFTR activity in the cell in thepresence of the candidate agent and the disclosed compound; and (iii)comparing the CFTR activity to that in the absence of the test agent,wherein an increase in CFTR activity in the presence of the test agentindicates that the agent increases CFTR activity. In certainembodiments, the cell expresses a mutant CFTR protein. In certainembodiments, CFTR activity is measured by measuring chloride channelactivity of the CFTR, and/or other ion transport activity. In certain ofthese embodiments, the method is high-throughput. In certain of theseembodiments, the candidate agent is a CFTR corrector or a CFTRpotentiator.

DETAILED DESCRIPTION

As used herein, the words “a” and “an” are meant to include one or moreunless otherwise specified. For example, the term “an agent” encompassesboth a single agent and a combination of two or more agents.

As discussed above, the present disclosure is directed in part tocompounds as described herein having the Formula (IV) or (V), or apharmaceutically acceptable salt, prodrug or solvate thereof,pharmaceutical compositions, methods of increasing CFTR activity andmethods of treating cystic fibrosis.

For example, disclosed herein, in an embodiment, are compounds havingthe Formula (IV) or (V):

and pharmaceutically acceptable salts, stereoisomers, and prodrugsthereof, wherein:

X₁ is CH or N; wherein when X₁ is N, R^(z) is not present and thenitrogen with R^(z) has a double bond with the adjacent N and there is asingle bond between X₁ and the adjacent N, and when X₁ is CH, X₁ has adouble bond with the nitrogen adjacent, and IV is selected from thegroup consisting of H, and C₁₋₃alkyl, and the nitrogen with IV has asingle bond with the adjacent N;

X₃ is selected from the group consisting of O, S, and NR_(hh);

pp is 1, 2, or 3;

R₁₁ is independently selected for each occurrence from the groupconsisting of hydrogen, halogen, C₁₋₄alkyl (optionally substituted byone, two or three halogens);

L₁ is selected from the group consisting of C₁₋₆ alkylene,C₃₋₆cycloalkylene, C₃₋₆cycloalkylene-C₁₋₄alkylene,C₁₋₃alkylene-NR_(hh)-S(O)_(w-), —C₁₋₃ alkylene-S(O)_(w)—NR_(hh)—,C₃₋₆cycloalkylene-C₀₋₂alkylene-S(O)_(w)—NR_(hh), andC₃₋₆cycloalkylene-C₀₋₂alkylene NR_(hh)—S(O)_(w-), wherein L₁ may beoptionally substituted by one, two or three substituents selected fromthe group consisting of halogen, hydroxyl, C₁-₃alkyl (optionallysubstituted by one, two or three substituents each selectedindependently from R_(ff));

R₄₄ is selected from the group consisting of H, halogen, hydroxyl,C₁₋₃alkoxy, heterocycle, and a 5-6 membered monocyclic or 8-10memberedbicyclic heteroaryl having one, two or three heteroatoms each selectedfrom O, N, and S; wherein the heterocycle and the heteroaryl may beoptionally substituted by one or two substituents each selectedindependently from R_(gg);

R_(ff) is selected for each occurrence from group consisting of halogen,hydroxyl, C₁₋₄alkyl, C₁₋₄alkyoxy, C₂₋₄alkenyl, C₃₋₆cycloalkyl, —NR′R″,—NR′—S(O)_(w)—C₁₋₃alkyl, S(O)_(w)—NR′R″, and —S(O)_(w)—C₁₋₃alkyl, wherew is 0, 1, or 2, wherein C₁₋₄alkyl, C₁₋₄alkyoxy, C₂₋₄alkenyl andC₃₋₆cycloalkyl may be optionally substituted by one, two or threesubstituents each independently selected from the group consisting ofhalogen, hydroxyl, —NR′R″, —NR′—S(O)_(w)—C₁₋₃alkyl, S(O)_(w)—NR′R″, and—S(O)_(w)—C₁₋₃alkyl;

R_(gg) is selected for each occurrence from group consisting of halogen,hydroxyl, C₁₋₆alkyl, C₁₋₆alkyoxy, C₂₋₆alkenyl, C₃₋₆cycloalkyl, —NR′R″,—NR′—S(O)_(w)—C₁₋₃alkyl, S(O)_(w)—NR′R″, and —S(O)_(w)—C₁₋₃alkyl, wherew is 0, 1, or 2, wherein C₁₋₆alkyl, C₁₋₆alkyoxy, C₂₋₆alkenyl andC₃₋₆cycloalkyl may each be optionally substituted by one, two or threesubstituents each independently selected from the group consisting ofhalogen, C₁₋₆alkyl, C₁₋₆alkoxy, hydroxyl, C(O)OH, —C(O)OC₁₋₆alkyl,—O—C₃₋₆cycloalkyl, —O-heterocycle, —O-heteroaryl, —O-phenyl, —NR′R″,—NR′—S(O)_(w)—C₁₋₃alkyl, S(O)_(w)—NR′R″, and —S(O)w-C₁₋₃alkyl;

w is 0, 1 or 2; and

R_(hh), is selected for each occurrence from the group consisting of H,C₁₋₆alkyl and C₃₋₆cycloalkyl.

In certain embodiments, L₁ may be C₁₋₃alkylene or C₃₋₅cycloalkyleneand/or X₁ is CH or C.

Also provided herein are compounds having a formula selected from

wherein qq is 0or 1 (and R₄₄, R₁₁ are defined above).

For example, provided herein are compounds having formula:

In certain embodiments, R₄₄ of one or more of above formulas is selectedfrom the group consisting of: pyrrolidinyl, piperidinyl,tetrahydropyranyl, and tetrahydrofuranyl. In other embodiments, R₄₄ isselected from the group consisting of:

wherein X independently for each occurrence is selected from the groupconsisting of O, S, NR_(hh), C C(R₈₈), and C(R₈₈)(R₉₉); X₂ independentlyfor each occurrence is selected from the group consisting of O, S andNR_(hh); R″ is H or C₁₋₄alkyl, each R₆₆, R₇₇, R₈₈and R₉₉ isindependently selected for each occurrence from H and R_(gg), and n is0, 1, 2, or 3.

In certain embodiments, each R₆₆, R₇₇, R₈₈and R₉₉ is independentlyselected for each occurrence from the group consisting of hydrogen,halogen, hydroxyl, C₁₋₆ alkyl, C₃₋₆ cycloalkyl, and heterocycle, whereinC₁₋₆ alkyl, C₃₋₆cycloalkyl, and heterocycle are optionally substitutedby one, two or three substituents each independently selected from thegroup consisting of hydroxyl, C₁₋₆ alkyl, C₁₋₆ alkoxy (optionallysubstituted by C₃₋₆cycloalkyl, heterocycle, —C₁₋₂alkyl-heterocycle andC₁₋₂alkyl-C₃₋₆cycloalkyl), —S(O)_(w)—C₁₋₃ alkyl (w is 0,1, or 2) and—NR′S(O)₂C₁₋₆ alkyl; and R′ is independently selected for eachoccurrence from H and C₁₋₄ alkyl and/or pp is 0, 1 or 2, and R₁₁ isselected from H, F, or methyl.

Also disclosed herein are compounds such as those having the Formula(IVa) or (Va):

or a pharmaceutically acceptable salt, prodrug or solvate thereof,wherein:

R₁ is selected from the group consisting of:

R₂ is selected from the group consisting of optionally substituted aryland optionally substituted heteroaryl;

R_(1a) is selected from the group consisting of hydrogen, optionallysubstituted C₁-C₁₀alkyl, optionally substituted C₂-C₁₀alkenyl,optionally substituted C₂-C₁₀alkynyl, optionally substitutedC₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl, optionallysubstituted aryl, halo, OR_(c), NR_(d)R_(d), C(O)OR_(c), NO₂, CN,C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d), NR_(d)C(O)R_(c),NR_(d)S(O)_(n)R_(c), N(R_(d))(COOR_(c)), NR_(d)C(O)C(O)R_(c),NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)NR_(d)R_(d), NR_(d)S(O)_(n)R_(c),S(O)_(n)R_(c), S(O)_(n)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c),optionally substituted heterocyclic and optionally substitutedheteroaryl;

R_(4a) is selected from the group consisting of hydrogen, optionallysubstituted C₁-C₁₀alkyl, optionally substituted C₂-C₁₀alkenyl,optionally substituted C₂-C₁₀alkynyl, optionally substitutedC₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl, optionallysubstituted aryl, halo, OR_(c), S(O)_(n)R_(c), NR_(d)R_(d), C(O)OR_(c),NO₂, CN, C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d), NR_(d)C(O)R_(c),NR_(d)S(O)R_(c), N(R_(d))(COOR_(c)), NR_(d)C(O)C(O)R_(c),NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)R_(d)R_(d), NR_(d)S(O)_(n)R_(c),S(O)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c), optionally substitutedheterocyclic and optionally substituted heteroaryl;

R_(4b) is selected from the group consisting of hydrogen, optionallysubstituted C₁-C₁₀alkyl, optionally substituted C₂-C₁₀alkenyl,optionally substituted C₂-C₁₀alkynyl, optionally substitutedC₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl, optionallysubstituted aryl, optionally substituted heterocyclic and optionallysubstituted heteroaryl;

R_(a) is selected from the group consisting of hydrogen, optionallysubstituted C₁-C₁₀alkyl, optionally substituted C₂-C₁₀alkenyl,optionally substituted C₂-C₁₀alkynyl, optionally substitutedC₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl, optionallysubstituted heterocyclic, optionally substituted aryl, optionallysubstituted heteroaryl, C(O)OR_(c), C(O)R_(c), C(O)C(O)R_(c) andS(O)_(n)R_(c);

or alternatively, R_(a) and the nitrogen atom to which it is attached istaken together with an adjacent C(R_(b1))(R_(b1)) or C(R_(b2))(R_(b2))to form an optionally substituted, 4- to 12-membered heterocyclic ringcontaining one or more ring nitrogen atoms, wherein said heterocyclicring optionally contains one or more ring heteroatoms selected fromoxygen and sulfur;

each R_(b1) and R_(b2) is independently selected from the groupconsisting of hydrogen, optionally substituted C₁-C₁₀alkyl, optionallysubstituted C₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl,optionally substituted C₃-C₁₂cycloalkyl, optionally substitutedC₃-C₁₂cycloalkenyl, optionally substituted heterocyclic, optionallysubstituted aryl, optionally substituted heteroaryl, halo, OR_(c),NR_(d)R_(d), C(O)OR_(c), NO₂, CN, C(O)R_(c), C(O)C(O)R_(c),C(O)NR_(d)R_(d), NR_(d)C(O)R_(c), NR_(d)S(O)_(n)R_(c),N(R_(d))(COOR_(c)), NR_(d)C(O)C(O)R_(c), NR_(d)C(O)NR_(d)R_(d),NR_(d)S(O)_(n)NR_(d)R_(d), NR_(d)S(O)_(n)R_(c), S(O)_(n)R_(c),S(O)_(n)NR_(d)R_(d), OC(O)OR_(c) and (C═NR_(d))R_(c); or alternatively,two geminal R_(b1) groups or two geminal R_(b2) groups and the carbon towhich they are attached are taken together to form a C(O) group, or yetalternatively, two geminal R_(b1) groups or two geminal R_(b2) groupsare taken together with the carbon atom to which they are attached toform a spiro C₃-C₁₂cycloalkyl, a spiro C₃-C₁₂cycloalkenyl, a spiroheterocyclic, a spiro aryl or spiro heteroaryl, each optionallysubstituted;

Y is selected from the group consisting of S(O)_(n.), NR_(d),NR_(d)S(O)_(n), NR_(d)S(O)_(n)NR_(d), NR_(d)C(O), NR_(d)C(O)O,NR_(d)C(O)C(O), NR_(d)C(O)NR_(d), S(O)_(n)NR_(d), and O;

each R_(c) is independently selected from the group consisting ofhydrogen, optionally substituted C₁-C₁₀alkyl, optionally substitutedC₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl, optionallysubstituted C₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl,optionally substituted heterocyclic, optionally substituted aryl andoptionally substituted heteroaryl;

each R_(d) is independently selected from the group consisting ofhydrogen, optionally substituted C₁-C₁₀alkyl, optionally substitutedC₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl, optionallysubstituted C₁-C₁₀alkoxy, optionally substituted C₃-C₁₂cycloalkyl,optionally substituted C₃-C₁₂cycloalkenyl, optionally substitutedheterocyclic, optionally substituted aryl and optionally substitutedheteroaryl; or two geminal R_(d) groups are taken together with thenitrogen atom to which they are attached to form an optionallysubstituted heterocyclic or an optionally substituted heteroaryl;

k is 0or 1;

m is 0, 1, 2, 3, 4, or 5;

each n is independently 0, 1 or 2.

In some embodiments, m is 0, 1 or 2 (e.g., m is 0or m is 1). In someembodiments, k is 0. In some embodiments, m is 0, 1 or 2 (e.g., m is 0orm is 1), and k is 0.

In some embodiments, R_(1a) is hydrogen.

In some embodiments, R_(a) is hydrogen or C₁-C₄ alkyl (optionallysubstituted by 1, 2 or 3 halogens). In certain embodiments, R_(a) ishydrogen.

In some embodiments, R_(b1) and R_(b2) are each independently selectedfrom the group consisting of hydrogen, hydroxyl, C₁₋₄alkoxy (optionallysubstituted by one, two or three substituents independently selectedfrom halogen and hydroxyl) and C₁-C₄ alkyl (optionally substituted byone, two or three substituents independently selected from halogen andhydroxyl). In certain embodiments, R_(b1) and R_(b2) for each occurrenceare hydrogen.

In some embodiments, R₂ is selected from the group consisting of phenyland a 5-6 membered heteroaryl having one or two heteroatoms eachselected from N, S, and O, wherein R₂ is optionally substituted by oneor two substituents each independently selected from the groupconsisting of halogen, and C₁-C₄ alkyl (optionally substituted by one,two or three halogens).

In certain embodiments, R₂ is phenyl. In other embodiments, R₂ is phenylis substituted with one or two R_(5,) wherein each R₅ is independentlyselected from the group consisting of optionally substitutedC₁-C₁₀alkyl, optionally substituted C₂-C₁₀alkenyl, optionallysubstituted C₂-C₁₀alkynyl, and halo. In still other embodiments, R₂ is apara-substituted phenyl.

In other embodiments, R₂ is selected from the group consisting of:optionally substituted thienyl, optionally substituted furanyl andoptionally substituted pyridinyl.

In some embodiments, R_(4a) is selected from the group consisting ofoptionally substituted C₁-C₆ alkyl, optionally substitutedC₃-C₇cycloalkyl, phenyl, OR_(c), C(O)OR_(c), C(O)R_(c), optionallysubstituted heterocycle and optionally substituted heteroaryl, whereinR_(c) is selected, independently for each occurrence, from the groupconsisting of H and C₁₋₆alkyl.

In certain embodiments, R_(4a) is heterocycle, or a 5-6 memberedmonocyclic or a 8-10membered bicyclic heteroaryl having one, two orthree heteroatoms selected from N, S or O, wherein the heterocycle orheteroaryl are optionally substituted by one, two or three substituentsindependently selected for each occurrence from the group consisting ofhalogen, C₁₋₆alkyl (optionally substituted by one, two or threesubstituents each independently selected from halogen and hydroxyl),C₁₋₆alkoxy (optionally substituted by one, two or three halogens),hydroxyl, and NR_(d)R_(d) wherein R_(d) is independently for eachoccurrence selected from H and C₁₋₄alkyl, or the two R_(d)s takentogether with the N to which they are attached form a heterocyclicring). For example, R_(4a) can be selected from the group consisting oftetrahydropyranyl, thiadiazolyl, tetrahydrofuranyl, and morpholinyl. Asanother example, R_(4a) can be a monocyclic heteroaryl containing one,two or three ring nitrogen atoms. As a further example, R_(4a) can beselected from the group consisting of furanyl, pyridinyl, pyrazinyl,pyrazolyl, imidazolyl, isoxazolyl, triazolyl, thiazolyl, oxadiazolyl,thiadiazolyl, thienyl, piperazinyl, and benzimidazolyl, each optionallysubstituted.

In certain embodiments, R_(4a) is selected from the group consisting of:

wherein each X is independently O, S or NR_(g);

each R_(g) is independently selected from the group consisting ofhydrogen, C₁-C₄ alkyl, C₃-C₆cycloalkyl;

each R₆, R₇and R₈is independently selected for each occurrence from thegroup consisting of hydrogen, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₁₆alkynyl, C₃-C₇cycloalkyl, C₃-C₇cycloalkenyl, phenyl, heterocycle,heteroaryl, halo, hydroxyl, carboxyl, OR_(c), NR_(d)R_(d), C(O)OR_(c),CN, C(O)R_(c), wherein the C₁₋₆alkyl, C₂-C₆ alkenyl, C₂-C₁₆ alkynyl,C₃-C₇cycloalkyl, C₃-C₇cycloalkenyl, phenyl, heterocycle, and heteroarylof R₆, R₇and R₈may each be optionally substituted by one, two or threesubstituents selected from halo, hydroxyl, C₁₋₆alkyl and C₁₋₆alkoxy;

R_(c) is C₁₋₄alkyl; and

R_(d) is independently for each occurrence selected from the groupconsisting of H and C₁₋₄alkyl, or the two R_(d)s taken together with theN to which they are attached form a heterocyclic ring.

In certain embodiments, R_(4a) is an optionally substitutedC₃-C₇cycloalkyl (e.g., optionally substituted cyclopropyl or anoptionally substituted cyclobutyl).

In certain of these embodiments, R_(4a) is substituted with asubstituent having the formula:

wherein each R_(h) is independently selected for each occurrence fromthe group consisting of hydrogen, halo, hydroxyl, C₁-C₆ alkyl, andC₃-C₆cycloalkyl, or two geminal R_(h) groups are independently takentogether with the carbon atom to which they are attached to form anoptionally substituted carbocyclic or heterocycle;

R₉ is selected from the group consisting of hydrogen, halo, CN,hydroxyl, methyl (optionally substituted by one, two or threesubstituents selected from halogen and hydroxyl), C₂-C₄ alkenyl, C₂-C₄alkynyl, C₃-C₆cycloalkyl, C₁₋₆alkoxy, NR_(d)R_(d), C(O)OR_(c), NO₂, CN,C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d), NR_(d)C(O)R_(c),NR_(d)S(O)_(n)R_(c), NR_(d)(COOR_(c)), NR_(d)C(O)C(O)R_(c),NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)NR_(d)R_(d), NR_(d)S(O)_(n)R_(c),S(O)_(n)R_(c), S(O)_(n)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c);

R_(c) is independently selected for each occurrence from the groupconsisting of H, C₁-C₆ alkyl, C₃₋₆cycloalkyl, heterocycle, andheteroaryl;

R_(d) is independently selected for each occurrence from H andC₁₋₄alkyl, or the two R_(d)s taken together with the N to which they areattached form a heterocyclic ring; and p is 0, 1, or 2.

For example, R_(4a) can be selected from the group consisting of:

wherein each R₁₀is independently selected from the group consisting ofhydrogen, optionally substituted C₁-C₆ alkyl, optionally substitutedC₂-C₆ alkenyl, optionally substituted C₂-C₆ alkynyl, optionallysubstituted C₃-C₆cycloalkyl, optionally substituted C₃-C₆cycloalkenyl,optionally substituted aryl, halo, OR_(c), NR_(d)R_(d), C(O)OR_(c), NO₂,CN, C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d), NR_(d)C(O)R_(c),NR_(d)S(O)_(n)R_(c), NR_(d)(COOR_(c)), NR_(d)C(O)C(O)R_(c),NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)NR_(d)R_(d), NR_(d)S(O)_(n)R_(c),S(O)_(n)R_(c), S(O)_(n)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c),optionally substituted heterocyclic and optionally substitutedheteroaryl; alternatively, two geminal R₁₀groups are taken together withthe carbon atom to which they are attached to form a spiroC₃-C₇cycloalkyl, a spiro C₃-C₇cycloalkenyl, a spiro heterocyclic, aspiro aryl or spiro heteroaryl, each optionally substituted; or yetalternatively, two vicinal R₁₀groups are taken together with the carbonatoms to which they are attached to form a fused, optionally substitutedcyclic group selected from the group consisting of C₄-C₈cycloalkyl,C₄-C₈cycloalkenyl, 4- to 8-membered heterocyclic, aryl and heteroaryl,each optionally substituted; or further alternatively, two R₁₀groupsattached to non-adjacent carbon atoms are taken together with the carbonatoms to which they are attached to form a bridged cyclic group selectedfrom the group consisting of C₃-C₈cycloalkyl, C₃-C₈cycloalkenyl, and 4-to 8-membered heterocyclic, each optionally substituted;

each R_(h) is independently selected from the group consisting ofhydrogen, halo, optionally substituted C₁-C₁₀alkyl, and optionallysubstituted C₃-C₆cycloalkyl, or two geminal R_(b) groups areindependently taken together with the carbon atom to which they areattached to form an optionally substituted heterocyclic or an optionallysubstituted heteroaryl;

R₉ is selected from the group consisting of hydrogen, optionallysubstituted C₁-C₁₀alkyl, optionally substituted C₂-C₁₀alkenyl,optionally substituted C₂-C₁₀alkynyl, optionally substitutedC₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl, optionallysubstituted aryl, halo, OR_(c), NR_(d)R_(d), C(O)OR_(c), NO₂, CN,C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d), NR_(d)C(O)R_(c),NR_(d)S(O)_(n)R_(c), NR_(d)(COOR_(c)), NR_(d)C(O)C(O)R_(c),NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)NR_(d)R_(d), NR_(d)S(O)_(n)R_(c),S(O)_(n)R_(c), S(O)_(n)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c),optionally substituted heterocyclic and optionally substitutedheteroaryl; and p is 0, 1, or 2.

In some embodiments, Y is S, S(O)₂ or S(O)₂NR_(d).

In some embodiments, R_(4b) is heterocycle or a 5-6 membered monocyclicor a 8-10membered bicyclic heteroaryl having one, two or threeheteroatoms selected from N, S or O, wherein the heterocycle orheteroaryl are optionally substituted by one, two or three substituentsindependently selected for each occurrence from the group consisting ofhalogen, C₁₋₆alkyl (optionally substituted by one, two or threesubstituents each independently selected from halogen and hydroxyl),C₁₋₆alkoxy (optionally substituted by one, two or three halogens),hydroxyl, and NR_(d)R_(d) wherein R_(d) is independently for eachoccurrence selected from H and C₁₋₄alkyl, or the two R_(d)s takentogether with the N to which they are attached form a heterocyclicring). For example, R_(4b) can be selected from the group consisting offuranyl, pyridinyl, pyrazinyl, pyrazolyl, imidazolyl, isoxazolyl,triazolyl, thiazolyl, oxadiazolyl, thiadiazolyl, thienyl, piperazinyl,and benzimidazolyl, each optionally substituted.

Exemplary compounds are shown below in Table 1:

TABLE 1 Com- pound No. IV-1

IV-2

IV-3

IV-4

IV-5

IV-6

V-1

V-2

V-3

V-5

V-6

V-7

Also contemplated herein are pharmaceutical compositions that include adisclosed compound such as those compounds having Formula (IVa), (Va),(IV), or (V) and a pharmaceutically acceptable carrier or excipient. Incertain embodiments, the compositions can include at least oneadditional CFTR modulator as described anywhere herein or at least twoadditional CFTR modulators, each independently as described anywhereherein.

It is to be understood that the specific embodiments described hereincan be taken in combination with other specific embodiments delineatedherein. For example, as discussed above, in some embodiments, R_(2a) isfluoro, and in some embodiments described above, A is an optionallysubstituted imidazolyl or pyrazolyl. The disclosure, in an embodiment,thus encompasses compound of Formula (IVa), (Va), (IV), or (V), whereinR_(2a) is fluoro and A is an optionally substituted imidazolyl orpyrazolyl.

The features and other details of the disclosure will now be moreparticularly described. Before further description of the presentinvention, certain terms employed in the specification, examples andappended claims are collected here. These definitions should be read inlight of the remainder of the disclosure and as understood by a personof skill in the art. Unless defined otherwise, all technical andscientific terms used herein have the same meaning as commonlyunderstood by a person of ordinary skill in the art.

It will be appreciated that the description of the present inventionherein should be construed in congruity with the laws and principals ofchemical bonding.

The term “alkyl”, as used herein, unless otherwise indicated, refers toboth branched and straight-chain saturated aliphatic hydrocarbon groupshaving the specified number of carbon atoms; for example, “C₁-C₁₀alkyl”denotes alkyl having 1 to 10carbon atoms, and straight or branchedhydrocarbons of 1-6, 1-4, or 1-3 carbon atoms, referred to herein asC₁₋₆alkyl, C₁₋₄alkyl, and C₁₋₃alkyl, respectively. Examples of alkylinclude, but are not limited to, methyl, ethyl, n-propyl, i-propyl,n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, n-hexyl, 2-methylbutyl,2-methylpentyl, 2-ethylbutyl, 3-methylpentyl, and 4-methylpentyl.

The term, “alkenyl”, as used herein, refers to both straight andbranched-chain moieties having the specified number of carbon atoms andhaving at least one carbon-carbon double bond. Exemplary alkenyl groupsinclude, but are not limited to, a straight or branched group of 2-6 or3-4 carbon atoms, referred to herein as C₂₋₆alkenyl, and C₃₋₄alkenyl,respectively. Exemplary alkenyl groups include, but are not limited to,vinyl, allyl, butenyl, pentenyl, etc.

The term, “alkynyl”, as used herein, refers to both straight andbranched-chain moieties having the specified number or carbon atoms andhaving at least one carbon-carbon triple bond.

The term “cycloalkyl,” as used herein, refers to saturated cyclic alkylmoieties having 3 or more carbon atoms, for example, 3-10, 3-6, or 4-6carbons, referred to herein as C₃₋₁₀cycloalkyl, C₃₋₆cycloalkyl orC₄₋₆cycloalkyl, respectively for example. Examples of cycloalkylinclude, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl and adamantyl.

The term “cycloalkenyl,” as used herein, refers to cyclic alkenylmoieties having 3 or more carbon atoms.

The term “cycloalkynyl,” as used herein, refers to cyclic alkynylmoieties having 5 or more carbon atoms.

“Alkylene” means a straight or branched, saturated aliphatic divalentradical having the number of carbons indicated. “Cycloalkylene” refersto a divalent radical of carbocyclic saturated hydrocarbon group havingthe number of carbons indicated.

The term “alkoxy” as used herein refers to a straight or branched alkylgroup attached to oxygen (alkyl-O—). Exemplary alkoxy groups include,but are not limited to, alkoxy groups of 1-6 or 2-6 carbon atoms,referred to herein as C₁₋₆alkoxy, and C₂₋₆alkoxy, respectively.Exemplary alkoxy groups include, but are not limited to methoxy, ethoxy,isopropoxy, etc.

The term “heterocyclic” or “heterocycle” encompasses heterocycloalkyl,heterocycloalkenyl, heterobicycloalkyl, heterobicycloalkenyl,heteropolycycloalkyl, heteropolycycloalkenyl, and the like unlessindicated otherwise. Heterocycloalkyl refers to cycloalkyl groupscontaining one or more heteroatoms (O, S, or N) within the ring.Heterocycloalkenyl as used herein refers to cycloalkenyl groupscontaining one or more heteroatoms (O, S or N) within the ring.Heterobicycloalkyl refers to bicycloalkyl groups containing one or moreheteroatoms (O, S or N) within a ring. Heterobicycloalkenyl as usedherein refers to bicycloalkenyl groups containing one or moreheteroatoms (O, S or N) within a ring. A heterocycle can refer to, forexample, a saturated or partially unsaturated 4- to 12or 4-10-memberedring structure, including bridged or fused rings, and whose ringstructures include one to three heteroatoms, such as nitrogen, oxygen,and sulfur. Where possible, heterocyclic rings may be linked to theadjacent radical through carbon or nitrogen. Examples of heterocyclicgroups include, but are not limited to, pyrrolidine, piperidine,morpholine, thiomorpholine, piperazine, oxetane, azetidine,tetrahydrofuran or dihydrofuran, etc.

Cycloalkyl, cycloalkenyl, and heterocyclic groups also include groupssimilar to those described above for each of these respectivecategories, but which are substituted with one or more oxo moieties.

The term “aryl”, as used herein, refers to mono- or polycyclic aromaticcarbocyclic ring systems. A polycyclic aryl is a polycyclic ring systemthat comprises at least one aromatic ring. Polycyclic aryls can comprisefused rings, covalently attached rings or a combination thereof. Theterm “aryl” embraces aromatic radicals, such as, phenyl, naphthyl,indenyl, tetrahydronaphthyl, and indanyl. An aryl group may besubstituted or unsubstituted. In some embodiments, the aryl is aC₄-C₁₀aryl. Examples of optionally substituted aryl are phenyl,substituted phenyl, naphthyl and substituted naphthyl.

The term “heteroaryl”, as used herein, refers to aromatic carbocyclicgroups containing one or more heteroatoms (O, S, or N) within a ring. Aheteroaryl group, unless indicated otherwise, can be monocyclic orpolycyclic. A heteroaryl group may additionally be substituted orunsubstituted. Heteroaryl groups can also include ring systemssubstituted with one or more oxo moieties. A polycyclic heteroaryl cancomprise fused rings, covalently attached rings or a combinationthereof. A polycyclic heteroaryl is a polycyclic ring system thatcomprises at least one aromatic ring containing one or more heteroatomswithin a ring. Examples of heteroaryl groups include, but are notlimited to, pyridinyl, pyridazinyl, imidazolyl, pyrimidinyl, pyrazolyl,triazolyl, pyrazinyl, quinolyl, isoquinolyl, tetrazolyl, furyl, thienyl,isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolinyl,isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, cinnolinyl,indazolyl, indolizinyl, phthalazinyl, triazinyl, isoindolyl, purinyl,oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,benzotriazolyl, benzothiazolyl, benzoxazolyl, quinazolinyl,quinoxalinyl, naphthyridinyl, dihydroquinolyl, tetrahydroquinolyl,dihydroisoquinolyl, tetrahydroisoquinolyl, benzofuryl, furopyridinyl,pyrolopyrimidinyl, thiazolopyridinyl, oxazolopyridinyl and azaindolyl.The foregoing heteroaryl groups may be C-attached or heteroatom-attached(where such is possible). For instance, a group derived from pyrrole maybe pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached). In someembodiments, the heteroaryl is 4- to 12-membered heteroaryl. In yetother embodiments, the heteroaryl is a mono or bicyclic 4- to10-membered heteroaryl.

The term “substituted” refers to substitution by independent replacementof one, two, or three or more of the hydrogen atoms with substituentsincluding, but not limited to, and unless indicated otherwise,—C₁-C₁₂alkyl, —C₂-C₁₂alkenyl, —C₂-C₁₂alkynyl, —C₃-C₁₂cycloalkyl,—C₃-C₁₂cycloalkenyl, C₃-C₁₂cycloalkynyl, -heterocyclic, —F, —Cl, —Br,—I, —OH, —NO₂, —N₃, —CN, —NH₂, oxo, thioxo, —NR_(x)R_(x), dialkylamino,-diarylamino, -diheteroarylamino, —OR_(x), —C(O)R_(y), —C(O)C(O)R_(y),—OCO₂R_(y), —OC(O)R_(y), OC(O)C(O)R_(y), —NHC(O)R_(y), —NHCO₂R_(y),—NHC(O)C(O)R_(y), NHC(S)NH₂, —NHC(S)NHR_(x), —NHC(NH)NH₂,—NHC(NH)NHR_(x), —NHC(NH)R_(x), —C(NH)NHR_(x), and (C═NR_(x))R_(x);—NR_(x)C(O)R_(x), —NR_(x)C(O)N(R_(x))₂, —NR_(x)CO₂R_(y),—NR_(x)C(O)C(O)R_(y), —NR_(x)C(S)NH₂, —NR_(x)C(S)NHR_(x),—NR_(x)C(NH)NH₂, —NR_(x)C(NH)NHR_(x), —NR_(x)C(NH)R_(x),—C(NR_(x))NHR_(x)—S(O)R_(y), —NHSO₂R_(x), —CH₂NH₂, —CH₂SO₂CH₃, -aryl,-arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl,—C₃-C₁₂-cycloalkyl, -polyalkoxyalkyl, -polyalkoxy, -methoxymethoxy,-methoxyethoxy, —SH, —S—R_(x), or -methylthiomethyl, wherein R_(x) isselected from the group consisting of hydrogen, —C₁-C₁₂alkyl,—C₂-C₁₂alkenyl, —C₂-C₁₂alkynyl, —C₃-C₁₂cycloalkyl, -aryl, -heteroaryland -heterocyclic and —R_(y) is selected from the group consisting ofhydrogen, —C₁-C₁₂alkyl, —C₂-C₁₂alkenyl, —C₂-C₁₂alkynyl,—C₃-C₁₂cycloalkyl, -aryl, -heteroaryl, -heterocyclic, —NH₂,—NH—C₁-C₁₂alkyl, —NH—C₂-C₁₂alkenyl, —NH—C₂-C₁₂-alkynyl,—NH—C₃-C₁₂cycloalkyl, —NH-aryl, —NH-heteroaryl and —NH-heterocyclic. Itis understood that the aryls, heteroaryls, alkyls, and the like can befurther substituted.

The terms “halo” or “halogen” as used herein refer to F, Cl, Br, or I.

The term “haloalkyl” as used herein refers to an alkyl group having 1 to(2n+1) substituent(s) independently selected from F, Cl, Br or I, wheren is the maximum number of carbon atoms in the alkyl group. It will beunderstood that haloalkyl is a specific example of an optionallysubstituted alkyl.

The terms “hydroxy” and “hydroxyl” as used herein refers to the radical—OH.

As will be understood by the skilled artisan, “H” is the symbol forhydrogen, “N” is the symbol for nitrogen, “S” is the symbol for sulfur,and “O” is the symbol for oxygen. “Me” is an abbreviation for methyl.

The compounds of the disclosure may contain one or more chiral centersand, therefore, exist as stereoisomers. The term “stereoisomers” whenused herein consist of all enantiomers or diastereomers. These compoundsmay be designated by the symbols “(+),” “(−), ” “R” or “S,” depending onthe configuration of substituents around the stereogenic carbon atom,but the skilled artisan will recognize that a structure may denote achiral center implicitly. The present disclosure encompasses variousstereoisomers of these compounds and mixtures thereof. Mixtures ofenantiomers or diastereomers may be designated “(±)” in nomenclature,but the skilled artisan will recognize that a structure may denote achiral center implicitly.

The compounds of the disclosure may contain one or more double bondsand, therefore, exist as geometric isomers resulting from thearrangement of substituents around a carbon-carbon double bond. Thesymbol ═ denotes a bond that may be a single, double or triple bond asdescribed herein. Substituents around a carbon-carbon double bond aredesignated as being in the “Z” or “F” configuration wherein the terms“Z” and “F” are used in accordance with IUPAC standards. Unlessotherwise specified, structures depicting double bonds encompass boththe “E” and “Z” isomers. Substituents around a carbon-carbon double bondalternatively can be referred to as “cis” or “trans,” where “cis”represents substituents on the same side of the double bond and “trans”represents substituents on opposite sides of the double bond.

Compounds of the disclosure may contain a carbocyclic or heterocyclicring and therefore, exist as geometric isomers resulting from thearrangement of substituents around the ring. The arrangement ofsubstituents around a carbocyclic or heterocyclic ring are designated asbeing in the “Z” or “E” configuration wherein the terms “Z” and “E” areused in accordance with IUPAC standards. Unless otherwise specified,structures depicting carbocyclic or heterocyclic rings encompass both“Z” and “E” isomers. Substituents around a carbocyclic or heterocyclicring may also be referred to as “cis” or “trans”, where the term “cis”represents substituents on the same side of the plane of the ring andthe term “trans” represents substituents on opposite sides of the planeof the ring. Mixtures of compounds wherein the substituents are disposedon both the same and opposite sides of plane of the ring are designated“cis/trans.”

Individual enantiomers and diasterisomers of compounds of the presentdisclosure can be prepared synthetically from commercially availablestarting materials that contain asymmetric or stereogenic centers, or bypreparation of racemic mixtures followed by resolution methods wellknown to those of ordinary skill in the art. These methods of resolutionare exemplified by (1) attachment of a mixture of enantiomers to achiral auxiliary, separation of the resulting mixture of diastereomersby recrystallization or chromatography and liberation of the opticallypure product from the auxiliary, (2) salt formation employing anoptically active resolving agent, (3) direct separation of the mixtureof optical enantiomers on chiral liquid chromatographic columns or (4)kinetic resolution using stereoselective chemical or enzymatic reagents.Racemic mixtures can also be resolved into their component enantiomersby well known methods, such as chiral-phase liquid chromatography orcrystallizing the compound in a chiral solvent. Stereoselectivesyntheses, a chemical or enzymatic reaction in which a single reactantforms an unequal mixture of stereoisomers during the creation of a newstereocenter or during the transformation of a pre-existing one, arewell known in the art. Stereoselective syntheses encompass both enantio-and diastereoselective transformations, and may involve the use ofchiral auxiliaries. For examples, see Carreira and Kvaerno, Classics inStereoselective Synthesis, Wiley-VCH: Weinheim, 2009. Where a particularcompound is described or depicted, it is intended to encompass thatchemical structure as well as tautomers of that structure.

The term “enantiomerically pure” means a stereomerically purecomposition of a compound. For example, a stereochemically purecomposition is a composition that is free or substantially free of otherstereoisomers of that compound. In another example, for a compoundhaving one chiral center, an enantiomerically pure composition of thecompound is free or substantially free of the other enantiomer. In yetanother example, for a compound having two chiral centers, anenantiomerically pure composition is free or substantially free of theother diastereomers.

Where a particular stereochemistry is described or depicted it isintended to mean that a particular enantiomer is present in excessrelative to the other enantiomer. A compound has an R-configuration at aspecific position when it is present in excess compared to the compoundhaving an S-configuration at that position. A compound has anS-configuration at a specific position when it is present in excesscompared to the compound having an R-configuration at that position.

The compounds disclosed herein can exist in solvated as well asunsolvated forms with pharmaceutically acceptable solvents such aswater, ethanol, and the like, and it is intended that disclosedcompounds include both solvated and unsolvated forms. In one embodiment,a disclosed compound is amorphous or, in another embodiment, a singlepolymorph. In another embodiment, a disclosed compound is a mixture ofpolymorphs. In another embodiment, a disclosed compound is in acrystalline form.

Isotopically labeled compounds are also contemplated herein, which areidentical to those recited herein, except that one or more atoms arereplaced by an atom having an atomic mass or mass number different fromthe atomic mass or mass number usually found in nature. Examples ofisotopes that can be incorporated into compounds of the inventioninclude isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus,sulfur, fluorine and chlorine, such as ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O,³¹P, ³²P, ³⁵S, ¹⁸F, and ³⁶Cl, respectively. For example, a disclosedcompound may have one or more H atom replaced with deuterium.

Certain isotopically-labeled disclosed compounds (e.g., those labeledwith ³H and ¹⁴C) are useful in compound and/or substrate tissuedistribution assays. Tritiated (i.e., ³H) and carbon-14 (i.e., ¹⁴C)isotopes are particularly preferred for their ease of preparation anddetectability. Further, substitution with heavier isotopes such asdeuterium (i.e., ²H) may afford certain therapeutic advantages resultingfrom greater metabolic stability (e.g., increased in vivo half-life orreduced dosage requirements) and hence may be preferred in somecircumstances. Isotopically labeled compounds can generally be preparedby following procedures analogous to those disclosed in the examplesherein by substituting an isotopically labeled reagent for anon-isotopically labeled reagent.

In some embodiments one or more of the nitrogen atoms of a disclosedcompound if present are oxidized to N-oxide.

Representative synthetic routes for the preparation of the compoundsdisclosed herein are provided throughout the Examples section. As willbe understood by the skilled artisan, diastereomers can be separatedfrom the reaction mixture using column chromatography.

Compounds of the invention can also be prepared using methods describedin the literature, including, but not limited to, J. Med. Chem. 2011,54(13), 4350-64; Russian Journal of Organic Chemistry, 2011, 47(8),1199-1203; U.S. Patent Application Publication No. 2009/0036451 A1;WO2008/046072 A2, and U.S. Pat. No. 4,336,264, the contents of each ofwhich are expressly incorporated by reference herein.

As discussed above, the invention encompasses a method of enhancing(e.g., increasing) CFTR activity in a subject (e.g., a subject sufferingfrom any one or more of the conditions described herein) comprisingadministering a compound of the invention in an effective amount. Theinvention also encompasses a method of treating a patient suffering froma condition associated with CFTR activity comprising administering tosaid patient an effective amount of a compound described herein. Incertain embodiments, the disease is cystic fibrosis.

“Treating” or “treatment” includes preventing or delaying the onset ofthe symptoms, complications, or biochemical indicia of a disease,alleviating or ameliorating the symptoms or arresting or inhibitingfurther development of the disease, condition, or disorder. A “subject”is an animal to be treated or in need of treatment. A “patient” is ahuman subject in need of treatment.

An “effective amount” refers to that amount of an agent that issufficient to achieve a desired and/or recited effect. In the context ofa method of treatment, an “effective amount” of the therapeutic agentthat is sufficient to ameliorate of one or more symptoms of a disorderand/or prevent advancement of a disorder, cause regression of thedisorder and/or to achieve a desired effect.

The term “modulating” encompasses increasing, enhancing, inhibiting,decreasing, suppressing, and the like. The terms “increasing” and“enhancing” mean to cause a net gain by either direct or indirect means.As used herein, the terms “inhibiting” and “decreasing” encompasscausing a net decrease by either direct or indirect means.

In some examples, CFTR activity is enhanced after administration of acompound described herein when there is an increase in the CFTR activityas compared to that in the absence of the administration of thecompound. CFTR activity encompasses, for example, chloride channelactivity of the CFTR, and/or other ion transport activity (for example,HCO₃ ⁻ transport). In certain of these embodiments, the activity of oneor more (e.g., one or two) mutant CFTRs (e.g., ΔF508, S549N, G542X,G551D, R117H, N1303K, W1282X, R553X, 621+1G>T, 1717−1G>A, 3849+10kbC>T,2789+5G>A, 3120+1G>A, I507de1, R1162X, 1898+1G>A, 3659de1C, G85E,D1152H, R560T, R347P, 2184insA, A455E, R334W, Q493X, and 2184delA CFTR)is enhanced (e.g., increased). Contemplated patients may have a CFTRmutation(s) from one or more classes, such as without limitation, ClassI CFTR mutations, Class II CFTR mutations, Class III CFTR mutations,Class IV CFTR mutations, Class V CFTR mutations, and Class VI mutations.Contemplated subject (e.g., human subject) CFTR genotypes include,without limitation, homozygote mutations (e.g., ΔF508/ΔF508andR117H/R117H) and compound heterozygote mutations (e.g., ΔF508/G551D;ΔF508/A455E; ΔF508/G542X; 4508F/W1204X; R553X/W1316X; W1282X/N1303K,591Δ18/E831X, F508del/R117H/N1303K/3849+10kbC>T; Δ303K/384; andDF508/G178R).

In certain embodiments, the mutation is a Class I mutation, e.g., aG542X; a Class II/I mutation, e.g., a ΔF508/G542X compound heterozygousmutation. In other embodiments, the mutation is a Class III mutation,e.g., a G551D; a Class II/Class III mutation, e.g., a ΔF508/G551Dcompound heterozygous mutation. In still other embodiments, the mutationis a Class V mutation, e.g., a A455E; Class II/Class V mutation, e.g., aΔF508/A455E compound heterozygous mutation. Of the more than 1000knownmutations of the CFTR gene, ΔF508is the most prevalent mutation of CFTRwhich results in misfolding of the protein and impaired trafficking fromthe endoplasmic reticulum to the apical membrane (Dormer et al. (2001).J Cell Sci 114, 4073-4081; http://www.genet.sickkids.on.ca/app). Incertain embodiments, ΔF508CFTR activity is enhanced (e.g., increased).In certain embodiments, ΔF508CFTR activity and/or G542X CFTR activityand/or G551D CFTR activity and/or A455E CFTR activity is enhanced (e.g.,increased). An enhancement of CFTR activity can be measured, forexample, using literature described methods, including for example,Ussing chamber assays, patch clamp assays, and hBE Ieq assay (Devor etal. (2000), Am J Physiol Cell Physiol 279(2): C461-79; Dousmanis et al.(2002), J Gen Physiol 119(6): 545-59; Bruscia et al. (2005), PNAS103(8): 2965-2971).

As discussed above, a method of treating cystic fibrosis is providedherein comprising administering a disclosed compound. Disclosed methodscan also be used to treat other conditions associated with CFTRactivity, including conditions associated with deficient CFTR activity.

In some embodiments, a method of treating a condition associated withdeficient or decreased CFTR activity is provided comprisingadministering an effective amount of a compound of Formula (IVa) or (Va)or (IV) or (V) that enhances CFTR activity. Non-limiting examples ofconditions associated with deficient CFTR activity are cystic fibrosis,congenital bilateral absence of vas deferens (CBAVD), acute, recurrent,or chronic pancreatitis, disseminated bronchiectasis, asthma, allergicpulmonary aspergillosis, smoking-related lung diseases, such as chronicobstructive pulmonary disease (COPD), chronic sinusitis, dry eyedisease, protein C deficiency, Aβ-lipoproteinemia, lysosomal storagedisease, type 1 chylomicronemia, mild pulmonary disease, lipidprocessing deficiencies, type 1 hereditary angioedema,coagulation-fibrinolyis, hereditary hemochromatosis, CFTR-relatedmetabolic syndrome, chronic bronchitis, constipation, pancreaticinsufficiency, hereditary emphysema, and Sjogren's syndrome.

In some embodiments, disclosed methods of treatment further compriseadministering an additional therapeutic agent. For example, in anembodiment, provided herein is a method of administering a disclosedcompound and at least one additional therapeutic agent. In certainaspects, a disclosed method of treatment comprises administering adisclosed compound, and at least two additional therapeutic agents.Additional therapeutic agents include, for example, mucolytic agents,bronchodilators, antibiotics, anti-infective agents, anti-inflammatoryagents, ion channel modulating agents, therapeutic agents used in genetherapy, CFTR correctors, and CFTR potentiators, or other agents thatmodulates CFTR activity. In some embodiments, at least one additionaltherapeutic agent is selected from the group consisting of a CFTRcorrector and a CFTR potentiator. Non-limiting examples of CFTRcorrectors and potentiators include VX-770(Ivacaftor), VX-809(3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)cyclopropanecarboxamido)-3-methylpyridin-2-yl)benzoicacid, VX-661(1-(2,2-difluoro-1,3-benzodioxol-5-yl)-N-[1-[(2R)-2,3-dihydroxypropyl]-6-fluoro-2-(2-hydroxy-1,1-dimethylethyl)-1H-indol-5-yl]-cyclopropanecarboxamide),VX-983, VX-152, VX-440, and Ataluren (PTC124)(3-[5-(2-fluorophenyl)-1,2,4-oxadiazol-3-yl]benzoic acid), FDL169,GLPG1837/ABBV-974 (for example, a CFTR potentiator), GLPG 2665, GLPG2222(for example, a CFTR corrector); and compounds described in, e.g.,WO2014/144860and 2014/176553, hereby incorporated by reference.Non-limiting examples of modulators include QBW-251, QR-010, NB-124, andcompounds described in, e.g., WO2014/045283; WO2014/081821,WO2014/081820, WO2014/152213; WO2014/160440, WO2014/160478,US2014027933; WO2014/0228376, WO2013/038390,WO2011/113894,WO2013/038386; and WO2014/180562, of which the disclosedmodulators in those publications are contemplated as an additionaltherapeutic agent and incorporated by reference. Non-limiting examplesof anti-inflammatory agents include N6022 (3-(5-(4-(1H-imidazol-1-yl)phenyl)-1-(4-carbamoyl-2-methylphenyl)-¹H-pyrrol-2-yl) propanoic acid),CTX-4430, N1861, N1785, and N91115.

In some embodiments, the methods described herein can further includeadministering an additional therapeutic agent or administering at leasttwo additional CFTR therapeutic agents. In some embodiments, the methodsdescribed herein can further include administering an additional CFTRmodulator or administering at least two additional CFTR modulators. Incertain embodiments, at least one CFTR modulator is a CFTR corrector(e.g., VX-809, VX-661, VX-983, VX-152, VX-440, GLPG2222 and GLPG2665) orpotentiator (e.g., ivacaftor, genistein and GLPG1837). In certain ofthese embodiments, one of the at least two additional therapeutic agentsis a CFTR corrector (e.g., VX-809, VX-661, VX-983, VX-152, and VX-440)and the other is a CFTR potentiator (e.g., ivacaftor and genistein). Incertain of these embodiments, one of the at least two additionaltherapeutic agents is a CFTR corrector (e.g., GLPG2222 or GLPG2665) andthe other is a CFTR potentiator (e.g., GLPG1837). In certain of theseembodiments, one of the at least two additional therapeutic agents is aCFTR corrector (e.g., VX-809 or VX-661) and the other is a CFTRpotentiator (e.g., ivacaftor). In certain of these embodiments, at leastone CFTR modulator is an agent that enhances read-through of stop codons(e.g., NB124 or ataluren).

Accordingly, in another aspect, this disclosure provides a method oftreating a condition associated with deficient or decreased CFTRactivity (e.g., cystic fibrosis), which includes administering to asubject in need thereof (e.g., a human patient in need thereof) aneffective amount of a disclosed compound and at least one or twoadditional CFTR therapeutic agent(s) (e.g., at least one or twoadditional CFTR therapeutic agents, e.g., in which one of the at leastone or two additional therapeutic agents is optionally a CFTR correctoror modulator (e.g., VX-809, VX-661, VX-983, VX-152, VX-440, GLPG2222,GLPG2665, NB124, ataluren) and/or the other is a CFTR potentiator (e.g.,ivacaftor, genistein, and GLPG1837); e.g., one of the at least twoadditional therapeutic agents is GLPG2222 or GLPG2665, and the other isGLPG1837; or one of the at least two additional therapeutic agents isVX-809 or VX-661, and the other is ivacaftor. In certain embodiments,the subject's CFTR genotype includes, without limitation, one or moreClass I CFTR mutations, one or more Class II CFTR mutations, one or moreClass III CFTR mutations, one or more Class IV CFTR mutations, or one ormore Class V CFTR mutations, or one or more Class VI CFTR mutations. Incertain embodiments, the subject's CFTR genotype includes, withoutlimitation, one or more homozygote mutations (e.g., ΔF508/ΔF508orR117H/R117H) and/or one or more compound heterozygote mutations (e.g.,ΔF508/G551D; ΔF508/A455E; ΔF508/G542X; Δ508F/W1204X; R553X/W1316X;W1282X/N1303K; F508del/R117H; N1303K/3849+10kbC>T; ΔF508/R334W;DF508/G178R. and 591Δ18/E831X). In certain embodiments, the subject'sCFTR genotype includes a Class I mutation, e.g., a G542X Class Imutation, e.g., a ΔF508/G542X compound heterozygous mutation. In otherembodiments, the subject's CFTR genotype includes a Class III mutation,e.g., a G551D Class III mutation, e.g., a ΔF508/G551D compoundheterozygous mutation. In still other embodiments, the subject's CFTRgenotype includes a Class V mutation, e.g., a A455E Class V mutation,e.g., a ΔF508/A455E compound heterozygous mutation. In certainembodiments, ΔF508CFTR activity and/or G542X CFTR activity and/or G551DCFTR activity and/or A455E activity is enhanced (e.g., increased). Incertain embodiments, the enhancement in activity (e.g., increase inactivity) provided by the combination of the disclosed compound and oneor two additional therapeutic agents is greater than additive whencompared to the enhancement in activity provided by each therapeuticcomponent individually.

Class Effect on CFTR protein Example of mutation I Shortened proteinW1282X Instead of inserting the amino acid tryptophan (W), the proteinsequence is prematurely stopped (indicated by an X). II Protein fails toreach cell ΔF508 A phenylalanine amino membrane acid (F) is deleted IIIChannel cannot be regulated G551D A “missense” mutation: properlyinstead of a glycine amino acid (G), aspartate (D) is added IV Reducedchloride conductance R117H Missense V Reduced due to incorrect 3120 + 1G > A Splice-site splicing of gene mutation in gene intron 16 VI Reduceddue to protein N287Y a A -> T at 991 instability

Genotype Description Possible Symptoms Δ508F/Δ508F homozygote Severelung disease, pancreatic insufficient R117H/R117H homozygote Congenitalbilateral absence of the vas deferens, No lung or pancreas disease,WT/Δ508F heterozygote Unaffected WT/3120 + 1 G > A heterozygoteUnaffected Δ508F/W1204X compound No lung disease, pancreaticheterozygote insufficient R553X and W1316X compound Mild lung disease,heterozygote pancreatic insufficient 591Δ18/E831X compound No lung orpancreas disease, heterozygote nasal polyps

For example, provided herein is a method of treating a patient havingone or more of the following mutations in the CFTR gene: G1244E, G1349D,G178R, G551S, S1251N, S1255P, S549N, S549R , G970R, or R117H, and/ore.g., a patient with one or two copies of the F508del mutation, or onecopy of the ΔF508mutation and a second mutation that results in a gatingeffect in the CFTR protein (e.g., a patient that is heterozygous forΔF508and G551D mutation), a patient with one copy of the ΔF508mutationand a second mutation that results in residual CFTR activity, or apatient with one copy of the ΔF508mutation and a second mutation thatresults in residual CFTR activity, comprising administering an effectiveamount of a disclosed compound. As described herein, such exemplarymethods (e.g., of a patient having one or mutations such as thosedescribed above) may include, for example, administering to such patienta combination therapy, e.g., administering (simultaneously orsequentially) an effective amount of ivacaftor to said patient and aneffective amount of disclosed compound that may act as an amplifier.Such administration may result, for example, in increased chloridetransport in human bronchial epithelial cells with e.g., one or twocopies of mutations, e.g, ΔF508mutation, as compared to administrationof ivacaftor alone. Another combination therapy that includes adisclosed compound may also include an effective amount of a readthroughagent (e.g., ataluren, NB124) and an effect amount of disclosed compoundthat may act as an amplifier.

The phrase “combination therapy,” as used herein, refers to anembodiment where a patient is co-administered a disclosed compound, aCFTR potentiator agent (e.g., ivacaftor) and optionally, one or moreCFTR corrector agent(s) (e.g, VX-661 and/or lumacaftor) as part of aspecific treatment regimen intended to provide the beneficial effectfrom the co-action of these therapeutic agents. For example, abeneficial effect of a combination may include, but is not limited to,pharmacokinetic or pharmacodynamic co-action resulting from thecombination of therapeutic agents. For example, administration of adisclosed compound with ivacaftor alone or with a CFTR corrector agent(e.g., lumacaftor or VX-661) may result in a level of function (e.g., asmeasured by chloride activity in HBE cells or patients that have aΔF508mutation, that achieves clinical improvement (or better) ascompared to the chloride activity level in cells or patients with aG551D mutation receiving ivacaftor alone, or ivacaftor and a correctoragent (lumacaftor or VX-661); or for example, administration of adisclosed compound with ivacaftor alone or ivacaftor with a CFTRcorrector agent (e.g., lumacaftor or VX-661) may result in a level offunction (e.g., as measured by chloride activity in HBE cells orpatients that have a A455E mutation, that achieves clinical improvement(or better) as compared to the chloride activity level at e.g., 50% ormore of wild type cells; or upon administration of a disclosed compoundand ivacaftor to a patient (e.g. having a G551D class III mutation) mayshow e.g., about two times or more improved activity of ivacaftor ascompared to administration of ivacaftor alone. Administration ofdisclosed therapeutic agents in combination typically is carried outover a defined time period (usually a day, days, weeks, months or yearsdepending upon the combination selected). Combination therapy isintended to embrace administration of multiple therapeutic agents in asequential manner, that is, wherein each therapeutic agent isadministered at a different time, as well as administration of thesetherapeutic agents, or at least two of the therapeutic agents, in asubstantially simultaneous manner. Substantially simultaneousadministration can be accomplished, for example, by administering to thesubject a single tablet or capsule having a fixed ratio of eachtherapeutic agent or in multiple, single capsules for each of thetherapeutic agents. Sequential or substantially simultaneousadministration of each therapeutic agent can be effected by anyappropriate route including, but not limited to, oral routes,inhalational routes, intravenous routes, intramuscular routes, anddirect absorption through mucous membrane tissues. The therapeuticagents can be administered by the same route or by different routes. Forexample, a first therapeutic agent of the combination selected may beadministered by intravenous injection or inhalation or nebulizer whilethe other therapeutic agents of the combination may be administeredorally.

Alternatively, for example, all therapeutic agents may be administeredorally or all therapeutic agents may be administered by intravenousinjection, inhalation or nebulization.

Combination therapy also can embrace the administration of thetherapeutic agents as described above in further combination with otherbiologically active ingredients and non-drug therapies. Where thecombination therapy further comprises a non-drug treatment, the non-drugtreatment may be conducted at any suitable time so long as a beneficialeffect from the co-action of the combination of the therapeutic agentsand non-drug treatment is achieved. For example, in appropriate cases,the beneficial effect is still achieved when the non-drug treatment istemporally removed from the administration of the therapeutic agents,perhaps by a day, days or even weeks.

The components of a disclosed combination may be administered to apatient simultaneously or sequentially. It will be appreciated that thecomponents may be present in the same pharmaceutically acceptablecarrier and, therefore, are administered simultaneously. Alternatively,the active ingredients may be present in separate pharmaceuticalcarriers, such as, conventional oral dosage forms, that can beadministered either simultaneously or sequentially.

In a further aspect, a method of identifying a candidate agent thatincreases CFTR activity is provided, which includes: (i) contacting acell that expresses a CFTR protein with the candidate agent and adisclosed compound; (ii) measuring the CFTR activity in the cell in thepresence of the candidate agent and the disclosed compound; and (iii)comparing the CFTR activity to that in the absence of the test agent,wherein an increase in CFTR activity in the presence of the test agentindicates that the agent increases CFTR activity. In certainembodiments, the cell expresses a mutant CFTR protein. In certainembodiments, CFTR activity is measured by measuring chloride channelactivity of the CFTR, and/or other ion transport activity. In certain ofthese embodiments, the method is high-throughput. In certain of theseembodiments, the candidate agent is a CFTR corrector or a CFTRpotentiator.

The term “pharmaceutically acceptable salt(s)” as used herein refers tosalts of acidic or basic groups that may be present in a disclosedcompounds used in disclosed compositions. Compounds included in thepresent compositions that are basic in nature are capable of forming awide variety of salts with various inorganic and organic acids. Theacids that may be used to prepare pharmaceutically acceptable acidaddition salts of such basic compounds are those that form non-toxicacid addition salts, i.e., salts containing pharmacologically acceptableanions, including, but not limited to, malate, oxalate, chloride,bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate,isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate,tannate, pantothenate, bitartrate, ascorbate, succinate, maleate,gentisinate, fumarate, gluconate, glucaronate, saccharate, formate,benzoate, glutamate, methanesulfonate, ethanesulfonate,benzenesulfonate, p-toluenesulfonate and pamoate (i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Compounds includedin the present compositions that are acidic in nature are capable offorming base salts with various pharmacologically acceptable cations.Examples of such salts include alkali metal or alkaline earth metalsalts, particularly calcium, magnesium, sodium, lithium, zinc,potassium, and iron salts. Compounds included in the presentcompositions that include a basic or acidic moiety may also formpharmaceutically acceptable salts with various amino acids. Thecompounds of the disclosure may contain both acidic and basic groups;for example, one amino and one carboxylic acid group. In such a case,the compound can exist as an acid addition salt, a zwitterion, or a basesalt.

Also contemplated herein are methods that include administering prodrugsof the compounds described herein, for example, prodrugs of a compoundof Formula (IVa) or (Va) or (IV) or (V), or a pharmaceutical compositionthereof or method of use of the prodrug.

The term “prodrug” refers to compounds that are transformed in vivo toyield a disclosed compound or a pharmaceutically acceptable salt,hydrate or solvate of the compound. The transformation may occur byvarious mechanisms (such as by esterase, amidase, phosphatase, oxidativeand or reductive metabolism) in various locations (such as in theintestinal lumen or upon transit of the intestine, blood or liver).Prodrugs are well known in the art (for example, see Rautio,Kumpulainen, et al, Nature Reviews Drug Discovery 2008, 7, 255). Forexample, if a compound of the invention or a pharmaceutically acceptablesalt, hydrate or solvate of the compound contains a carboxylic acidfunctional group, a prodrug can comprise an ester formed by thereplacement of the hydrogen atom of the acid group with a group such as(C₁₋₈)alkyl, (C₂₋₁₂)alkylcarbonyloxymethyl, 1-(alkylcarbonyloxy)ethylhaving from 4 to 9 carbon atoms, 1-methyl-1-(alkylcarbonyloxy)-ethylhaving from 5 to 10carbon atoms, alkoxycarbonyloxymethyl having from 3to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7carbonatoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to 8carbonatoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9 carbon atoms,1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10carbon atoms,3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl,di-N,N-(C₁₋₂)alkylamino(C₂₋₃)alkyl (such as β-dimethylaminoethyl),carbamoyl-(C₁₋₂)alkyl, N,N-di(C₁₋₂)alkylcarbamoyl-(C₁₋₂)alkyl andpiperidino-, pyrrolidino- or morpholino(C₂₋₃)alkyl.

Similarly, if a compound of the invention contains an alcohol functionalgroup, a prodrug can be formed by the replacement of the hydrogen atomof the alcohol group with a group such as (C₁₋₆)alkylcarbonyloxymethyl,1-((C₁₋₆)alkylcarbonyloxy)ethyl, 1-methyl-1((C₁₋₆)alkylcarbonyloxy)ethyl(C₁₋₆)alkoxycarbonyloxymethyl, N-(C₁₋₆)alkoxycarbonylaminomethyl,succinoyl, (C₁₋₆)alkylcarbonyl, α-amino(C₁₋₄)alkylcarbonyl,arylalkylcarbonyl and α-aminoalkylcarbonyl, orα-aminoalkylcarbonyl-α-aminoalkylcarbonyl, where eachα-aminoalkylcarbonyl group is independently selected from the naturallyoccurring L-amino acids, P(O)(OH)₂, —P(O)(O(C₁₋₆)alkyl)₂or glycosyl (theradical resulting from the removal of a hydroxyl group of the hemiacetalform of a carbohydrate).

If a compound of the invention incorporates an amine functional group, aprodrug can be formed, for example, by creation of an amide orcarbamate, an N-alkylcarbonyloxyalkyl derivative, an(oxodioxolenyl)methyl derivative, an N-Mannich base, imine or enamine Inaddition, a secondary amine can be metabolically cleaved to generate abioactive primary amine, or a tertiary amine can metabolically cleavedto generate a bioactive primary or secondary amine. For examples, seeSimplício, et al., Molecules 2008, 13, 519 and references therein

Also contemplated in certain embodiments is the use of clathrates of thecompounds described herein, pharmaceutical compositions comprising theclathrates, and methods of use of the clathrates. Clathrates of adisclosed compound or a pharmaceutical composition thereof are alsocontemplated herein.

As discussed above, the disclosure also contemplates administration ofpharmaceutical compositions comprising a pharmaceutically acceptablecarrier or excipient and a compound described herein. A disclosedcompound, or a pharmaceutically acceptable salt, solvate, clathrate orprodrug therof, can be administered in pharmaceutical compositionscomprising a pharmaceutically acceptable carrier or excipient. Theexcipient can be chosen based on the expected route of administration ofthe composition in therapeutic applications. The route of administrationof the composition depends on the condition to be treated. For example,intravenous injection may be preferred for treatment of a systemicdisorder and oral administration may be preferred to treat agastrointestinal disorder. The route of administration and the dosage ofthe composition to be administered can be determined by the skilledartisan without undue experimentation in conjunction with standarddose-response studies. Relevant circumstances to be considered in makingthose determinations include the condition or conditions to be treated,the choice of composition to be administered, the age, weight, andresponse of the individual patient, and the severity of the patient'ssymptoms. A pharmaceutical composition comprising a disclosed compoundor a pharmaceutically acceptable salt, solvate, clathrate or prodrug,can be administered by a variety of routes including, but not limitedto, parenteral, oral, pulmonary, ophthalmic, nasal, rectal, vaginal,aural, topical, buccal, transdermal, intravenous, intramuscular,subcutaneous, intradermal, intraocular, intracerebral, intralymphatic,intraarticular, intrathecal and intraperitoneal. The compositions canalso include, depending on the formulation desired,pharmaceutically-acceptable, non-toxic carriers or diluents, which aredefined as vehicles commonly used to formulate pharmaceuticalcompositions for animal or human administration. The diluent is selectedso as not to affect the biological activity of the pharmacologic agentor composition. Examples of such diluents are distilled water,physiological phosphate-buffered saline, Ringer's solutions, dextrosesolution, and Hank's solution. In addition, the pharmaceuticalcomposition or formulation may also include other carriers, adjuvants,or nontoxic, nontherapeutic, nonimmunogenic stabilizers and the like.Pharmaceutical compositions can also include large, slowly metabolizedmacromolecules such as proteins, polysaccharides such as chitosan,polylactic acids, polyglycolic acids and copolymers (such as latexfunctionalized SEPHAROSE™, agarose, cellulose, and the like), polymericamino acids, amino acid copolymers, and lipid aggregates (such as oildroplets or liposomes).

Disclosed compositions can be administered parenterally such as, forexample, by intravenous, intramuscular, intrathecal or subcutaneousinjection. Parenteral administration can be accomplished byincorporating a composition into a solution or suspension. Suchsolutions or suspensions may also include sterile diluents such as waterfor injection, saline solution, fixed oils, polyethylene glycols,glycerine, propylene glycol or other synthetic solvents. Parenteralformulations may also include antibacterial agents such as, for example,benzyl alcohol or methyl parabens, antioxidants such as, for example,ascorbic acid or sodium bisulfite and chelating agents such as EDTA.Buffers such as acetates, citrates or phosphates and agents for theadjustment of tonicity such as sodium chloride or dextrose may also beadded. The parenteral preparation can be enclosed in ampules, disposablesyringes or multiple dose vials made of glass or plastic. Additionally,auxiliary substances, such as wetting or emulsifying agents,surfactants, pH buffering substances and the like can be present incompositions. Other components of pharmaceutical compositions are thoseof petroleum, animal, vegetable, or synthetic origin, for example,peanut oil, soybean oil, and mineral oil. In general, glycols such aspropylene glycol or polyethylene glycol are preferred liquid carriers,particularly for injectable solutions.

Injectable formulations can be prepared either as liquid solutions orsuspensions; solid forms suitable for solution in, or suspension in,liquid vehicles prior to injection can also be prepared. The preparationalso can also be emulsified or encapsulated in liposomes or microparticles such as polylactide, polyglycolide, or copolymer for enhancedadjuvant effect, as discussed above [Langer, Science 249: 1527, 1990andHanes, Advanced Drug Delivery Reviews 28: 97-119, 1997]. Thecompositions and pharmacologic agents described herein can beadministered in the form of a depot injection or implant preparationwhich can be formulated in such a manner as to permit a sustained orpulsatile release of the active ingredient.

Additional formulations suitable for other modes of administrationinclude oral, intranasal, and pulmonary formulations, suppositories,transdermal applications and ocular delivery. For suppositories, bindersand carriers include, for example, polyalkylene glycols ortriglycerides; such suppositories can be formed from mixtures containingthe active ingredient in the range of about 0.5% to about 10%,preferably about 1% to about 2%. Oral formulations include excipients,such as pharmaceutical grades of mannitol, lactose, starch, magnesiumstearate, sodium saccharine, cellulose, and magnesium carbonate. Topicalapplication can result in transdermal or intradermal delivery.Transdermal delivery can be achieved using a skin patch or usingtransferosomes. [Paul et al., Eur. J. Immunol. 25: 3521-24, 1995; Cevcet al., Biochem. Biophys. Acta 1368: 201-15, 1998].

For the purpose of oral therapeutic administration, the pharmaceuticalcompositions can be incorporated with excipients and used in the form oftablets, troches, capsules, elixirs, suspensions, syrups, wafers,chewing gums and the like. Tablets, pills, capsules, troches and thelike may also contain binders, excipients, disintegrating agent,lubricants, glidants, sweetening agents, and flavoring agents. Someexamples of binders include microcrystalline cellulose, gum tragacanthor gelatin. Examples of excipients include starch or lactose. Someexamples of disintegrating agents include alginic acid, corn starch andthe like. Examples of lubricants include magnesium stearate or potassiumstearate. An example of a glidant is colloidal silicon dioxide. Someexamples of sweetening agents include sucrose, saccharin and the like.Examples of flavoring agents include peppermint, methyl salicylate,orange flavoring and the like. Materials used in preparing these variouscompositions should be pharmaceutically pure and non-toxic in theamounts used. In another embodiment, the composition is administered asa tablet or a capsule.

Various other materials may be present as coatings or to modify thephysical form of the dosage unit. For instance, tablets may be coatedwith shellac, sugar or both. A syrup or elixir may contain, in additionto the active ingredient, sucrose as a sweetening agent, methyl andpropylparabens as preservatives, a dye and a flavoring such as cherry ororange flavor, and the like. For vaginal administration, apharmaceutical composition may be presented as pessaries, tampons,creams, gels, pastes, foams or spray.

The pharmaceutical composition can also be administered by nasaladministration. As used herein, nasally administering or nasaladministration includes administering the composition to the mucusmembranes of the nasal passage or nasal cavity of the patient. As usedherein, pharmaceutical compositions for nasal administration of acomposition include therapeutically effective amounts of the compoundsprepared by well-known methods to be administered, for example, as anasal spray, nasal drop, suspension, gel, ointment, cream or powder.Administration of the composition may also take place using a nasaltampon or nasal sponge.

For topical administration, suitable formulations may includebiocompatible oil, wax, gel, powder, polymer, or other liquid or solidcarriers. Such formulations may be administered by applying directly toaffected tissues, for example, a liquid formulation to treat infectionof conjunctival tissue can be administered dropwise to the subject'seye, or a cream formulation can be administered to the skin.

Rectal administration includes administering the pharmaceuticalcompositions into the rectum or large intestine. This can beaccomplished using suppositories or enemas. Suppository formulations caneasily be made by methods known in the art. For example, suppositoryformulations can be prepared by heating glycerin to about 120° C.,dissolving the pharmaceutical composition in the glycerin, mixing theheated glycerin after which purified water may be added, and pouring thehot mixture into a suppository mold.

Transdermal administration includes percutaneous absorption of thecomposition through the skin. Transdermal formulations include patches,ointments, creams, gels, salves and the like.

In addition to the usual meaning of administering the formulationsdescribed herein to any part, tissue or organ whose primary function isgas exchange with the external environment, for purposes of the presentinvention, “pulmonary” will also mean to include a tissue or cavity thatis contingent to the respiratory tract, in particular, the sinuses. Forpulmonary administration, an aerosol formulation containing the activeagent, a manual pump spray, nebulizer or pressurized metered-doseinhaler as well as dry powder formulations are contemplated. Suitableformulations of this type can also include other agents, such asantistatic agents, to maintain the disclosed compounds as effectiveaerosols.

A drug delivery device for delivering aerosols comprises a suitableaerosol canister with a metering valve containing a pharmaceuticalaerosol formulation as described and an actuator housing adapted to holdthe canister and allow for drug delivery. The canister in the drugdelivery device has a head space representing greater than about 15% ofthe total volume of the canister. Often, the compound intended forpulmonary administration is dissolved, suspended or emulsified in amixture of a solvent, surfactant and propellant. The mixture ismaintained under pressure in a canister that has been sealed with ametering valve.

The disclosure also encompasses the treatment of a condition associatedwith a dysfunction in proteostasis in a subject comprising administeringto said subject an effective amount of a disclosed compound thatenhances, improves or restores proteostasis of a protein. Proteostasisrefers to protein homeostasis. Dysfunction in protein homeostasis is aresult of protein misfolding, protein aggregation, defective proteintrafficking or protein degradation. For example, the disclosurecontemplates administering a disclosed compound e.g., Formula (IVa),(Va), (IV), or (V) that corrects protein misfolding, reduces proteinaggregation, corrects or restores protein trafficking and/or affectsprotein degradation for the treatment of a condition associated with adysfunction in proteostasis. In some aspects, a disclosed compound e.g.,Formula (IVa), (Va), (IV), or (V) that corrects protein misfoldingand/or corrects or restores protein trafficking is administered. Incystic fibrosis, the mutated or defective enzyme is the cystic fibrosistransmembrane conductance regulator (CFTR). One of the most commonmutations of this protein is ΔF508which is a deletion (Δ) of threenucleotides resulting in a loss of the amino acid phenylalanine (F) atthe 508th (508) position on the protein. As described above, mutatedcystic fibrosis transmembrane conductance regulator exists in amisfolded state and is characterized by altered trafficking as comparedto the wild type CFTR. Additional exemplary proteins of which there canbe a dysfunction in proteostasis, for example that can exist in amisfolded state, include, but are not limited to, glucocerebrosidase,hexosamine A, aspartylglucsaminidase, α-galactosidase A, cysteinetransporter, acid ceremidase, acid α-L-fucosidase, protective protein,cathepsin A, acid β-glucosidase, acid β-galactosidase, iduronate2-sulfatase, α-L-iduronidase, galactocerebrosidase, acid α-mannosidase,acid β-mannosidase, arylsulfatase B, arylsulfatase A,N-acetylgalactosamine-6-sulfate sulfatase, acid β-galactosidase,N-acetylglucosamine-1-phosphotransferase, acid sphingmyelinase, NPC-1,acid α-glucosidase, β-hexosamine B, heparin N-sulfatase,α-N-acetylglucosaminidase, α-glucosaminide N-acetyltransferase,N-acetylglucosamine-6-sulfate sulfatase, α-N-acetylgalactosaminidase,α-neuramidase, β-glucuronidase, β-hexosamine A and acid lipase,polyglutamine, α-synuclein, TDP-43, superoxide dismutase (SOD), Aβpeptide, tau protein, transthyretin and insulin. The compounds ofFormula (IVa), (Va), (IV), or (V) can be used to restore proteostasis(e.g., correct folding and/or alter trafficking) of the proteinsdescribed above.

Protein conformational diseases encompass gain of function disorders andloss of function disorders. In one embodiment, the proteinconformational disease is a gain of function disorder. The terms “gainof function disorder,” “gain of function disease,” “gain of toxicfunction disorder” and “gain of toxic function disease” are usedinterchangeably herein. A gain of function disorder is a diseasecharacterized by increased aggregation-associated proteotoxicity. Inthese diseases, aggregation exceeds clearance inside and/or outside ofthe cell. Gain of function diseases include, but are not limited to,neurodegenerative diseases associated with aggregation of polyglutamine,Lewy body diseases, amyotrophic lateral sclerosis,transthyretin-associated aggregation diseases, Alzheimer's disease,Machado-Joseph disease, cerebral B-amyloid angiopathy, retinal ganglioncell degeneration, tauopathies (progressive supranuclear palsy,corticobasal degeneration, frontotemporal lobar degeneration), cerebralhemorrhage with amyloidosis, Alexander disease, Serpinopathies, familialamyloidotic neuropathy, senile systemic amyloidosis, ApoAI amyloidosis,ApoAII amyloidosis, ApoAIV amyloidosis, familial amyloidosis of theFinnish type, lysozyme amyloidosis, fibrinogen amyloidosis, dialysisamyloidosis, inclusion body myositis/myopathy, cataracts, medullarythyroid carcinoma, cardiac atrial amyloidosis, pituitary prolactinoma,hereditary lattice corneal dystrophy, cutaneous lichen amyloidosis,corneal lactoferrin amyloidosis, corneal lactoferrin amyloidosis,pulmonary alveolar proteinosis, odontogenic tumor amyloid, seminalvesical amyloid, sickle cell disease, critical illness myopathy, vonHippel-Lindau disease, spinocerebellar ataxia 1, Angelman syndrome,giant axon neuropathy, inclusion body myopathy with Paget disease ofbone, frontotemporal dementia (IBMPFD) and prion diseases.Neurodegenerative diseases associated with aggregation of polyglutamineinclude, but are not limited to, Huntington's disease, dentatorubral andpallidoluysian atrophy, several forms of spino-cerebellar ataxia, andspinal and bulbar muscular atrophy Alzheimer's disease is characterizedby the formation of two types of aggregates: extracellular aggregates ofAβ peptide and intracellular aggregates of the microtubule associatedprotein tau. Transthyretin-associated aggregation diseases include, forexample, senile systemic amyloidoses and familial amyloidoticneuropathy. Lewy body diseases are characterized by an aggregation ofα-synuclein protein and include, for example, Parkinson's disease, Lewybody dementia (LBD) and multiple system atrophy (SMA). Prion diseases(also known as transmissible spongiform encephalopathies or TSEs) arecharacterized by aggregation of prion proteins. Exemplary human priondiseases are Creutzfeldt-Jakob Disease (CJD), Variant Creutzfeldt-JakobDisease, Gerstmann-Straussler-Scheinker Syndrome, Fatal FamilialInsomnia and Kuru. In another embodiment, the misfolded protein isalpha-1 anti-trypsin.

In a further embodiment, the protein conformation disease is a loss offunction disorder. The terms “loss of function disease” and “loss offunction disorder” are used interchangeably herein. Loss of functiondiseases are a group of diseases characterized by inefficient folding ofa protein resulting in excessive degradation of the protein. Loss offunction diseases include, for example, lysosomal storage diseases.Lysosomal storage diseases are a group of diseases characterized by aspecific lysosomal enzyme deficiency which may occur in a variety oftissues, resulting in the build-up of molecules normally degraded by thedeficient enzyme. The lysosomal enzyme deficiency can be in a lysosomalhydrolase or a protein involved in the lysosomal trafficking. Lysosomalstorage diseases include, but are not limited to,aspartylglucosaminuria, Fabry's disease, Batten disease, Cystinosis,Farber, Fucosidosis, Galactasidosialidosis, Gaucher's disease (includingTypes 1, 2 and 3), Gm1 gangliosidosis, Hunter's disease, Hurler-Scheie'sdisease, Krabbe's disease, α-Mannosidosis, β-Mannosidosis,Maroteaux-Lamy's disease, Metachromatic Leukodystrophy, Morquio Asyndrome, Morquio B syndrome, Mucolipidosis II, Mucolipidosis III,Neimann-Pick Disease (including Types A, B and C), Pompe's disease,Sandhoff disease, Sanfilippo syndrome (including Types A, B, C and D),Schindler disease, Schindler-Kanzaki disease, Sialidosis, Sly syndrome,Tay-Sach's disease and Wolman disease.

In another embodiment, the disease associated with a dysfunction inproteostasis is a cardiovascular disease. Cardiovascular diseasesinclude, but are not limited to, coronary artery disease, myocardialinfarction, stroke, restenosis and arteriosclerosis. Conditionsassociated with a dysfunction of proteostasis also include ischemicconditions, such as, ischemia/reperfusion injury, myocardial ischemia,stable angina, unstable angina, stroke, ischemic heart disease andcerebral ischemia.

In yet another embodiment, the disease associated with a dysfunction inproteostasis is diabetes and/or complications of diabetes, including,but not limited to, diabetic retinopathy, cardiomyopathy, neuropathy,nephropathy, and impaired wound healing.

In a further embodiment, the disease associated with a dysfunction inproteostasis is an ocular disease including, but not limited to,age-related macular degeneration (AMD), diabetic macular edema (DME),diabetic retinopathy, glaucoma, cataracts, retinitis pigmentosa (RP) anddry macular degeneration.

In yet additional embodiments, a disclosed method is directed totreating a disease associated with a dysfunction in proteostasis,wherein the disease affects the respiratory system or the pancreas. Incertain additional embodiments, a contemplated method encompass treatinga condition selected from the group consisting ofpolyendocrinopathy/hyperinsulinemia, diabetes mellitus, Charcot-MarieTooth syndrome, Pelizaeus-Merzbacher disease, and Gorham's Syndrome.

Additional conditions associated with a dysfunction of proteostasisinclude hemoglobinopathies, inflammatory diseases, intermediate filamentdiseases, drug-induced lung damage and hearing loss. For example,provided herein are methods for the treatment of hemoglobinopathies(such as sickle cell anemia), an inflammatory disease (such asinflammatory bowel disease, colitis, ankylosing spondylitis),intermediate filament diseases (such as non-alcoholic and alcoholicfatty liver disease) and drug induced lung damage (such asmethotrexate-induced lung damage). In another embodiment, methods fortreating hearing loss, such as noise-induced hearing loss,aminoglycoside-induced hearing loss, and cisplatin-induced hearing losscomprising administering a disclosed compound are provided.

Additional contemplated conditions include those associated with adefect in protein trafficking and that can be treated according todisclosed methods include: PGP mutations, hERG trafficking mutations,nephrongenic diabetes insipidus mutations in the arginine-vasopressinreceptor 2, persistent hyperinsulinemic hypoglycemia of infancy (PHH1)mutations in the sulfonylurea receptor 1, and α1AT.

The invention is illustrated by the following examples which are notmeant to be limiting in any way.

EXEMPLIFICATION

The compounds described herein can be prepared in a number of ways basedon the teachings contained herein and synthetic procedures known in theart. In the description of the synthetic methods described below, it isto be understood that all proposed reaction conditions, including choiceof solvent, reaction atmosphere, reaction temperature, duration of theexperiment and workup procedures, can be chosen to be the conditionsstandard for that reaction, unless otherwise indicated. It is understoodby one skilled in the art of organic synthesis that the functionalitypresent on various portions of the molecule should be compatible withthe reagents and reactions proposed. Substituents not compatible withthe reaction conditions will be apparent to one skilled in the art, andalternate methods are therefore indicated. The starting materials forthe examples are either commercially available or are readily preparedby standard methods from known materials. At least some of the compoundsidentified as “intermediates” herein are contemplated as compounds ofthe invention.

Example 1 N-(2-methoxyethyl)-5-phenyl-1H-pyrazole-3-carboxamide

A solution of 5-phenyl-1H-pyrazole-3-carboxylic acid (0.15 g, 0.79 mmol)in THF (10mL), 2-methoxy-ethylamine (0.09 g, 1.18mmol), EDC.HCl (0.304g, 1.59 mmol) and HOBt (0.214 g, 1.59 mmol) was stirred for 18h at roomtemperature. Volatiles were removed under vacuum, poured in ice-water(50mL) and extracted with ethyl acetate (2×50mL). The combined organiclayer was dried over anhydrous Na₂SO₄ and concentrated under reducedpressure to get a brown liquid. The crude residue was purified bycombiflash using 20% MeOH in DCM to give the product (0.15 g, 78.9%) asoff white solid; ¹H NMR (400MHz, CDCl₃): δ 12.01 (br, 1H), 8.80(br, 1H),7.61-7.59 (d, J=7.4 Hz, 2H), 7.46-7.43 (t, 2H), 7.39-7.36 (t, 1H),7.10(s, 1H), 3.75-3.73 (m, 2H), 3.70-3.63 (m, 2H), 3.46 (s, 3H); LC-MS:[M+H]⁺246.2; HPLC purity 99.82% at 220nm and 99.82% at 254 nm.

Using above method the following compounds were synthesized usingrespective amine

Example 25-phenyl-N-((tetrahydrofuran-2-yl)methyl)-1H-pyrazole-3-carboxamide

Yield (0.14 g,63.6%) as off white solid; ¹H NMR (400MHz, CDCl₃): δ 12.19(br, 1H), 8.56 (br, 1H), 7.62-7.60(d, J=7.4 Hz, 2H), 7.46-7.42 (t, 2H),7.38-7.35 (t, 1H), 7.08(s, 1H), 4.22-4.20(m, 1H), 3.99-3.93 (m, 1H),3.86-3.81 (m, 1H), 3.76-3.70(m, 1H), 3.59-3.54 (m, 1H), 2.11-2.04 (m,1H), 1.98-1.91 (m, 2H), 1.75-1.70(m, 1H); LC-MS: [M+H]⁺271.8; HPLCpurity 99.05% at 220nm and 98.60% at 254 nm.

Example 3 N-(2-morpholinoethyl)-5-phenyl-1H-pyrazole-3-carboxamide

Yield (0.15 g,62.50%) as off white solid; ¹H NMR (400MHz, CDCl₃): δ10.89 (br, 1H), 7.62-7.60(d, J=7.4 Hz, 2H), 7.45-7.42 (t, 2H), 7.39-7.35(t,1H), 7.26 (br, 1H), 7.00(s, 1H), 3.72-3.70(m, 4H), 3.59-3.54 (m, 2H),2.61-2.58(t, 2H), 2.50(m, 4H); LC-MS: [M+H]⁺301.2; HPLC purity 99.68% at220nm and 99.66% at 254 nm.

Example 4N-(3-(1H-imidazol-1-yl)propyl)-5-phenyl-1H-pyrazole-3-carboxamide

Yield (0.145 g,57.31%) as off white solid; ¹H NMR (400MHz, CDCl₃): δ7.64-7.62 (d, J=7.32 Hz, 3H), 7.45-7.42 (t, 2H), 7.39-7.35 (t, 1H), 7.09(s, 1H), 7.05 (s, 1H), 6.98(s, 1H), 4.08-4.05 (m, 2H), 3.47-342 (m, 2H),2.16-2.09 (m, 2H); LC-MS: [M+H]⁺296.2; HPLC purity 99.85% at 220nm and99.75% at 254 nm.

Example 5 CFTR Activity Assays

i. Ussing Measurements

As discussed above, Ussing measurements are used to measure CFTRactivity. In this method, primary lung epithelial cells (hBEs)homozygous for the Cystic Fibrosis-causing ΔF508mutation aredifferentiated for a minimum of 4 weeks in an air-liquid interface onSnapWell filter plates prior to the Ussing measurements. Cells areapically mucus-washed for 30minutes prior to treatment with compounds.The basolateral media is removed and replaced with media containing thecompound of interest diluted to its final concentration from DMSOstocks. Treated cells are incubated at 37° C. and 5% CO₂ for 24 hours.At the end of the treatment period, the cells on filters are transferredto the Ussing chamber and equilibrated for 30minutes. The short-circuitcurrent is measured in voltage clamp-mode (V_(hold)=0mV), and the entireassay is conducted at a temperature of 36° C.-36.5° C. Once the voltagesstabilized, the chambers are clamped, and data is recorded by pulsereadings every 5 seconds. Following baseline current stabilization, thefollowing additions can be applied and the changes in current andresistance of the cells can be monitored:

-   -   1. Benzamil to the apical chamber to inhibit ENaC sodium channel    -   2. Forskolin to both chambers to activate ΔF508-CFTR by        phosphorylation.    -   3. Genistein to both chambers to potentiate ΔF508-CFTR channel        opening.    -   4. CFTRinh-172 to the apical chamber to inhibit ΔF508-CFTR        Cl-conductance.

The inhibitable current (that current that is blocked by CFTRinh-172) ismeasured as the specific activity of the ΔF508-CFTR channel, andincreases in response to compound in this activity over that observed invehicle-treated samples are identified as the correction of ΔF508-CFTRfunction imparted by the compound tested.

ii. hBE Equivalent Current (Ieq) Assay

Primary lung epithelial cells homozygous for the Cystic Fibrosis-causingΔF508mutation were differentiated for a minimum of 4 weeks in anair-liquid interface on Costar 24 well HTS filter plates prior to theequivalent current (Ieq) measurements. Cells were apically mucus-washedfor 30minutes 24 h prior to treatment with compounds. The basolateralmedia was removed and replaced with media containing the compound ofinterest diluted to its final concentration from DMSO stocks. Treatedcells were incubated at 37° C. and 5% CO₂ for 24 hours. At the end ofthe treatment period, the media was changed to the Ieq experimentalsolution for 30minutes before the experiment and plates are maintainedin a CO₂-free incubator during this period. The plates containing thecells were then placed in pre-warmed heating blocks at 36° C.±0.5 for 15minutes before measurements are taken. The transepithelial voltage(V_(T)) and conductance (G_(T)) were measured using a custom 24 channelcurrent clamp (TECC-24) with 24 well electrode manifold. The Ieq assaymeasurements were made following additions with standardized timeperiods:

-   -   1. The baseline V_(T) and G_(T) values were measured for        approximately 20minutes.    -   2. Benzamil was added to block ENaC for 15 minutes.    -   3. Forskolin plus VX-770were added to maximally activate        ΔF508-CFTR for 27minutes.    -   4. Bumetanide was added to inhibit the NaK₂Cl cotransporter and        shut-off secretion of chloride.

The activity data captured was the area under the curve (AUC) for thetraces of the equivalent chloride current. The AUC was collected fromthe time of the forskolin/VX-770addition until the inhibition bybumetanide addition. Correction in response to compound treatment wasscored as the increase in the AUC for compound-treated samples over thatof vehicle-treated samples. The results are shown below in Table 2. Theresults are shown below in Table 2. (** indicates activity ≥200% ofVX-809 (1 uM) with compound at 10uM and VX-809 at 1 uM; * indicatesactivity 100-200% of VX-809 (1 uM) with compound at 10uM and VX-809 at 1uM. ^(##) indicates activity≥200% of VX-809 (3 uM) with compound at 10uMand VX-809 at 3 uM; ^(#) indicates activity 100-200% of VX-809 (3 uM)with compound at 10uM and VX-809 at 3 uM.

TABLE 2 Structure Ieq

*

*

*

*

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

Example 6

i. Ussing Measurements

As discussed above, Ussing measurements can be used to measure CFTRactivity. In this method, primary lung epithelial cells (hBEs) with aCystic fibrosis causing class I mutation are differentiated for aminimum of 4 weeks in an air-liquid interface on SnapWell™ filter platesprior to the Ussing measurements. Cells are apically mucus-washed for30minutes prior to treatment with compounds. The basolateral media isremoved and replaced with media containing the compound of interestdiluted to its final concentration from DMSO or aqueous stocks. Treatedcells are incubated at 37° C. and 5% CO₂for 24 hours. At the end of thetreatment period, the cells on filters are transferred to the Ussingchamber and equilibrated for 30minutes. The short-circuit current ismeasured in voltage clamp-mode (V_(hold)=0mV), and the entire assay isconducted at a temperature of 36° C.-36.5° C. Once the voltagesstabilize, the chambers are clamped, and data are recorded by pulsereadings every 5 seconds. Following baseline current stabilization, thefollowing additions are applied and the changes in current andresistance of the cells are monitored:

-   -   1. Benzamil to the apical chamber to inhibit ENaC sodium        channel.    -   2. Forskolin to both chambers to activate ΔF508-CFTR by        phosphorylation.    -   3. Ivacaftor or Genistein to the apical chamber to potentiate        ΔF508-CFTR channel opening.    -   4. CFTRinh-172 to the apical chamber to inhibit ΔF508-CFTR        Cl-conductance.

The forskolin-sensitive current and inhibitable current (thatpotentiated current that is blocked by CFTRinh-172) are measured as thespecific activity of the ΔF508-CFTR channel, and increase in response tocompound in this activity over that observed in vehicle-treated samplesare identified as the correction of ΔF508-CFTR function imparted by thecompound tested.

Example 7

i. Ussing Measurements

As discussed above, Ussing measurements can be used to measure CFTRactivity. In this method, primary lung epithelial cells (hBEs) with aCystic Fibrosis-causing class III mutation are differentiated for aminimum of 4 weeks in an air-liquid interface on SnapWell™ filter platesprior to the Ussing measurements. Cells are apically mucus-washed for30minutes prior to treatment with compounds. The basolateral media isremoved and replaced with media containing the compound of interestdiluted to its final concentration from DMSO stocks. Treated cells areincubated at 37° C. and 5% CO₂ for 24 hours. At the end of the treatmentperiod, the cells on filters are transferred to the Ussing chamber andequilibrated for 30minutes. The short-circuit current is measured involtage clamp-mode (V_(hold)=0mV), and the entire assay is conducted ata temperature of 36° C.-36.5° C. Once the voltages stabilize, thechambers are clamped, and data is recorded by pulse readings every 5seconds. Following baseline current stabilization, the followingadditions are applied and the changes in current and resistance of thecells is monitored:

-   -   1. Benzamil to the apical chamber to inhibit ENaC sodium        channel.    -   2. Forskolin to both chambers to activate ΔF508-CFTR by        phosphorylation.    -   3. VX-770or Genistein to the apical chamber to potentiate        ΔF508-CFTR channel opening.    -   4. CFTRinh-172 to the apical chamber to inhibit ΔF508-CFTR        Cl-conductance.

The forskolin-sensitive current and inhibitable current (thatpotentiated current that is blocked by CFTRinh-172) are measured as thespecific activity of the ΔF508-CFTR channel, and increase in response tocompound in this activity over that observed in vehicle-treated samplesare identified as the correction of ΔF508-CFTR function imparted by thecompound tested.

Example 8

i. Ussing Measurements

As discussed above, Ussing measurements can be used to measure CFTRactivity. In this method, primary lung epithelial cells (hBEs) with aCystic Fibrosis-causing class V mutation are differentiated for aminimum of 4 weeks in an air-liquid interface on SnapWell™ filter platesprior to the Ussing measurements. Cells are apically mucus-washed for30minutes prior to treatment with compounds. The basolateral media isremoved and replaced with media containing the compound of interestdiluted to its final concentration from DMSO stocks. Treated cells areincubated at 37° C. and 5% CO₂ for 24 hours. At the end of the treatmentperiod, the cells on filters are transferred to the Us sing chamber andequilibrated for 30minutes. The short-circuit current is measured involtage clamp-mode (V_(hold)=0mV), and the entire assay is conducted ata temperature of 36° C.-36.5° C. Once the voltages stabilize, thechambers are clamped, and data is recorded by pulse readings every 5seconds. Following baseline current stabilization, the followingadditions are applied and the changes in current and resistance of thecells is monitored:

-   -   1. Benzamil to the apical chamber to inhibit ENaC sodium        channel.    -   2. Forskolin to both chambers to activate ΔF508-CFTR by        phosphorylation.    -   3. VX-770or Genistiein to the apical chamber to potentiate        ΔF508-CFTR channel opening.    -   4. CFTRinh-172 to the apical chamber to inhibit ΔF508-CFTR        Cl-conductance.

The forskolin-sensitive current and inhibitable current (thatpotentiated current that is blocked by CFTRinh-172) are measured as thespecific activity of the ΔF508-CFTR channel, and increases in responseto compound in this activity over that observed in vehicle-treatedsamples are identified as the correction of ΔF508-CFTR function impartedby the compound tested.

ii. JIBE Equivalent Current (Ieq) Assay

Primary lung epithelial cells homozygous for the Cystic Fibrosis-causingΔF508mutation are differentiated for a minimum of 4 weeks in anair-liquid interface on Costar 24 well HTS filter plates prior to theequivalent current (Ieq) measurements. Cells are apically mucus-washedfor 30minutes 24 h prior to treatment with compounds. The basolateralmedia is removed and replaced with media containing the compound ofinterest diluted to its final concentration from DMSO stocks. Treatedcells are incubated at 37° C. and 5% CO₂for 24 hours. At the end of thetreatment period, the media is changed to the Ieq experimental solutionfor 30minutes before the experiment and plates are maintained in aCO₂-free incubator during this period. The plates containing the cellsare then placed in pre-warmed heating blocks at 36° C±0.5 for 15 minutesbefore measurements are taken. The transepithelial voltage (V_(T)) andconductance (G_(T)) are measured using a custom 24 channel current clamp(TECC-24) with 24 well electrode manifold. The Ieq assay measurementsare made following additions with standardized time periods:

-   -   1. The baseline V_(T) and G_(T) values are measured for        approximately 20minutes.    -   2. Benzamil is added to block ENaC for 15 minutes.    -   3. Forskolin plus VX-770(ivacaftor) are added to maximally        activate ΔF508-CFTR for 27minutes.    -   4. Bumetanide is added to inhibit the NaK₂Cl cotransporter and        shut-off secretion of chloride.

The activity data captured is the area under the curve (AUC) for thetraces of the equivalent chloride current. The AUC is collected from thetime of the forskolin/VX-770addition until the inhibition by bumetanideaddition. Correction in response to compound treatment is scored as theincrease in the AUC for compound-treated samples over that ofvehicle-treated samples.

INCORPORATION BY REFERENCE

All publications and patents mentioned herein, including those itemslisted below, are hereby incorporated by reference in their entirety forall purposes as if each individual publication or patent wasspecifically and individually incorporated by reference. In case ofconflict, the present application, including any definitions herein,will control.

EQUIVALENTS

While specific embodiments of the subject invention have been discussed,the above specification is illustrative and not restrictive. Manyvariations of the invention will become apparent to those skilled in theart upon review of this specification. The full scope of the inventionshould be determined by reference to the claims, along with their fullscope of equivalents, and the specification, along with such variations.

Unless otherwise indicated, all numbers expressing quantities ofingredients, reaction conditions, and so forth used in the specificationand claims are to be understood as being modified in all instances bythe term “about.” Accordingly, unless indicated to the contrary, thenumerical parameters set forth in this specification and attached claimsare approximations that may vary depending upon the desired propertiessought to be obtained by the present invention.

1. A compound represented by formula IV or V:

and pharmaceutically acceptable salts, stereoisomers, and prodrugsthereof, wherein: X₁ is CH or N; wherein when X₁ is N, R^(z) is notpresent and the nitrogen with R^(z) has a double bond with the adjacentN and there is a single bond between X₁ and the adjacent N, and when X₁is CH, X₁ has a double bond with the nitrogen adjacent, and R^(z) isselected from the group consisting of H, and C₁₋₃alkyl, and the nitrogenwith R^(z) has a single bond with the adjacent N; X₃ is selected fromthe group consisting of O, S, and NR_(hh); pp for each occurrence isselected from the group consisting of 1, 2, or 3; Ru is independentlyselected for each occurrence from the group consisting of hydrogen,halogen, and C₁₋₄alkyl (optionally substituted by one, two or threehalogens); L₁ is selected from the group consisting of C₁₋₆alkylene,C₃₋₆cycloalkylene, C₃₋₆cycloalkylene-C₁₋₄alkylene,C₁₋₃alkylene-NR_(hh)—S(O)_(w)—, —C₁₋₃alkylene-S(O)_(w)—NR_(hh)—,C₃₋₆cycloalkylene-C₀₋₂alkylene-S(O)_(w)—NR_(hh), andC₃₋₆cycloalkylene-C₀₋₂alkylene NR_(hh)—S(O)_(w)—, wherein L₁ may beoptionally substituted by one, two or three substituents selected fromthe group consisting of halogen, hydroxyl, and C₁₋₃alkyl (optionallysubstituted by one, two or three substituents each selectedindependently from R_(ff)); R₄₄ is selected from the group consisting ofH, halogen, hydroxyl, C₁₋₃alkoxy, heterocycle, and a 5-6 memberedmonocyclic or 8-10membered bicyclic heteroaryl having one, two or threeheteroatoms each selected from O, N, and S; wherein the heterocycle andthe heteroaryl may be optionally substituted by one or two substituentseach selected independently from R_(gg); R_(ff) is selected for eachoccurrence from group consisting of halogen, hydroxyl, C₁₋₄alkyl,C₁₋₄alkyoxy, C₂₋₄alkenyl, C₃₋₆cycloalkyl, —NR′R″, S(O)_(w)—NR′R″, and—S(O)_(w)—C₁₋₃alkyl, where w is 0, 1, or 2, wherein C₁₋₄alkyl,C₁₋₄alkyoxy, C₂₋₄alkenyl and C₃₋₆cycloalkyl may be optionallysubstituted by one, two or three substituents each independentlyselected from the group consisting of halogen, hydroxyl, —NR′R″,S(O)_(w)—NR′R″, and —S(O)_(w)—C₁₋₃alkyl; R_(gg) is selected for eachoccurrence from group consisting of halogen, hydroxyl, C₁₋₆alkyl,C₁₋₆alkyoxy, C₂₋₆alkenyl, C₃₋₆cycloalkyl, S(O)_(w)—NR′R″, and—S(O)_(w)—C₁₋₃alkyl, where w is 0, 1, or 2, wherein C₁₋₆alkyl,C₁₋₆alkyoxy, C₂₋₆alkenyl and C₃₋₆cycloalkyl may each be optionallysubstituted by one, two or three substituents each independentlyselected from the group consisting of halogen, C₁₋₆alkyl, C₁₋₆alkoxy,hydroxyl, C(O)OH, —C(O)OC₁₋₆alkyl, —O—C₃₋₆cycloalkyl, —O-heterocycle,—O-heteroaryl, —O-phenyl, —NR′R″, S(O)_(w)—NR′R″, and—S(O)_(w)—C₁₋₃alkyl; w is 0, 1 or 2; and R_(hh) is selected for eachoccurrence from the group consisting of H, C₁₋₆alkyl and C₃₋₆cycloalkyl.2. The compound of claim 1, wherein L₁ is C₁₋₃alkylene orC₃₋₅cycloalkylene.
 3. The compound of claim 1, represented by:

wherein qq is 0or
 1. 4. The compound of claim 1, represented by:


5. The compound of claim 1, wherein R₄₄ is selected from the groupconsisting of pyrrolidinyl, piperidinyl, tetrahydropyranyl, andtetrahydrofuranyl.
 6. The compound of claim 1, wherein R₄₄ is selectedfrom the group consisting of:

wherein X independently for each occurrence is selected from the groupconsisting of O, S, NR_(hh), C, C(R₈₈), and _(c)(R₈₈)(R₉₉)_(;) X₂independently for each occurrence is selected from the group consistingof O, S and NR_(hh); R″ is H or C₁₋₄alkyl, each R₆₆, R₇₇, R₈₈and R₉₉ isindependently selected for each occurrence from H and R_(gg), and n is0, 1, 2, or
 3. 7. The compound of claim 6, wherein each R₆₆, R₇₇, R₈₈andR₉₉ is independently selected for each occurrence from the groupconsisting of hydrogen, halogen, hydroxyl, C₁₋₆alkyl, C₃₋₆cycloalkyl,and heterocycle, wherein C₁-₆alkyl, C₃₋₆cycloalkyl, and heterocycle areoptionally substituted by one, two or three substituents eachindependently selected from the group consisting of hydroxyl, C₁₋₆alkyl,C₁₋₆alkoxy (optionally substituted by C₃₋₆cycloalkyl, heterocycle,—C₁₋₂alkyl-heterocycle and C₁₋₂alkyl-C₃₋₆cycloalkyl) , —S(O)_(w)—C₁₋₃alkyl (w is 0, 1, or 2) and —NR′S(O)₂C₁₋₆ alkyl; and R′ is independentlyselected for each occurrence from H and C₁₋₄alkyl.
 8. The compound ofclaim 1 any one of claims 1 7, wherein pp is 0, 1 or 2, and R₁₁ isselected from H, F, or methyl.
 9. A compound having the Formula (IVa) orthe Formula (Va):

or a pharmaceutically acceptable salt, prodrug or solvate thereof,wherein: R₁ is selected from the group consisting of:

R₂ is selected from the group consisting of optionally substituted aryland optionally substituted heteroaryl; R_(3a) is selected from the groupconsisting of hydrogen, optionally substituted C₁-C₁₀alkyl, optionallysubstituted C₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl,optionally substituted C₃-C₁₂cycloalkyl, optionally substitutedC₃-C₁₂cycloalkenyl, optionally substituted aryl, halo, OR_(c),NR_(d)R_(d), C(O)OR_(c), NO₂, CN, C(O)R_(c), C(O)C(O)R_(c),C(O)NR_(d)R_(d), NR_(d)C(O)R_(c), NR_(d)S(O)_(n)R_(c),N(R_(d))(COOR_(c)), NR_(d)C(O)C(O)R_(c), NR_(d)C(O)NR_(d)R_(d),NR_(d)S(O)_(n)NR_(d)R_(d), NR_(d)S(O)_(n)R_(c), S(O)_(n)R_(c),S(O)_(n)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c), optionallysubstituted heterocyclic and optionally substituted heteroaryl; R_(4a)is selected from the group consisting of hydrogen, optionallysubstituted C₁-C₁₀alkyl, optionally substituted C₂-C₁₀alkenyl,optionally substituted C₂-C₁₀alkynyl, optionally substitutedC₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl, optionallysubstituted aryl, halo, OR_(c), S(O)_(n)R_(c), NR_(d)R_(d), C(O)OR_(c),NO₂, CN, C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d), NR_(d)C(O)R_(c),NR_(d)S(O)R_(c), N(R_(d))(COOR_(c)), NR_(d)C(O)C(O)R_(c),NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)R_(d)R_(d), NR_(d)S(O)_(n)R_(c),S(O)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c), optionally substitutedheterocyclic and optionally substituted heteroaryl; R_(4b) is selectedfrom the group consisting of hydrogen, optionally substitutedC₁-C₁₀alkyl, optionally substituted C₂-C₁₀alkenyl, optionallysubstituted C₂-C₁₀alkynyl, optionally substituted C₃-C₁₂cycloalkyl,optionally substituted C₃-C₁₂cycloalkenyl, optionally substituted aryl,optionally substituted heterocyclic and optionally substitutedheteroaryl; R_(a) is selected from the group consisting of hydrogen,optionally substituted C₁-C₁₀alkyl, optionally substitutedC₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl, optionallysubstituted C₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl,optionally substituted heterocyclic, optionally substituted aryl,optionally substituted heteroaryl, C(O)OR_(c), C(O)R_(c), C(O)C(O)R_(c)and S(O)_(n)R_(c); or alternatively, R_(a) and the nitrogen atom towhich it is attached is taken together with an adjacentC(R_(b1))(R_(b1)) or C(R_(b2))(R_(b2)) to form an optionallysubstituted, 4- to 12-membered heterocyclic ring containing one or morering nitrogen atoms, wherein said heterocyclic ring optionally containsone or more ring heteroatoms selected from oxygen and sulfur; eachR_(b1) and R_(b2) is independently selected from the group consisting ofhydrogen, optionally substituted C₁-C₁₀alkyl, optionally substitutedC₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl, optionallysubstituted C₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl,optionally substituted heterocyclic, optionally substituted aryl,optionally substituted heteroaryl, halo, OR_(c), NR_(d)R_(d),C(O)OR_(c), NO₂, CN, C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d),NR_(d)C(O)R_(c), NR_(d)S(O)_(n)R_(c), N(R_(d))(COOR_(c)),NR_(d)C(O)C(O)R_(c), NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)NR_(d)R_(d),NR_(d)S(O)_(n)R_(c), S(O)_(n)R_(c), S(O)_(n)NR_(d)R_(d), OC(O)OR_(c) and(C═NR_(d))R_(c); or alternatively, two geminal R_(b1) groups or twogeminal R_(b2) groups and the carbon to which they are attached aretaken together to form a C(O) group, or yet alternatively, two geminalR_(b1) groups or two geminal R_(b2) groups are taken together with thecarbon atom to which they are attached to form a spiro C₃-C₁₂cycloalkyl,a spiro C₃-C₁₂cycloalkenyl, a spiro heterocyclic, a spiro aryl or spiroheteroaryl, each optionally substituted; Y is selected from the groupconsisting of S(O)_(n,), NR_(d), NR_(d)S(O)_(n), NR_(d)S(O)_(n)NR_(d),NR_(d)C(O), NR_(d)C(O)O, NR_(d)C(O)C(O), NR_(d)C(O)NR_(d),S(O)_(n)NR_(d), and O; each R_(c) is independently selected from thegroup consisting of hydrogen, optionally substituted C₁-C₁₀alkyl,optionally substituted C₂-C₁₀alkenyl, optionally substitutedC₂-C₁₀alkynyl, optionally substituted C₃-C₁₂cycloalkyl, optionallysubstituted C₃-C₁₂cycloalkenyl, optionally substituted heterocyclic,optionally substituted aryl and optionally substituted heteroaryl; eachR_(d) is independently selected from the group consisting of hydrogen,optionally substituted C₁-C₁₀alkyl, optionally substitutedC₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl, optionallysubstituted C₁-C₁₀alkoxy, optionally substituted C₃-C₁₂cycloalkyl,optionally substituted C₃-C₁₂cycloalkenyl, optionally substitutedheterocyclic, optionally substituted aryl and optionally substitutedheteroaryl; or two geminal R_(d) groups are taken together with thenitrogen atom to which they are attached to form an optionallysubstituted heterocyclic or an optionally substituted heteroaryl; k is0or 1; m is 0, 1, 2, 3, 4, or 5; each n is independently 0, 1 or
 2. 10.The compound of claim 9, wherein m is 0, 1 or
 2. 11. The compound ofclaim 9, wherein m is
 0. 12. The compound of claim 9, wherein m is 1.13. The compound of claim 9, wherein R_(1a) is hydrogen.
 14. Thecompound of claim 9, wherein R_(a) is hydrogen or C₁-C₄ alkyl(optionally substituted by 1, 2 or 3 halogens).
 15. The compound ofclaim 9, wherein R_(a) is hydrogen.
 16. The compound of claim 9, whereineach of R_(b1) and R_(b2) is independently selected from of hydrogen,hydroxyl, C₁₋₄alkoxy (optionally substituted by one, two or threesubstituents independently selected from halogen and hydroxyl) and C₁-C₄alkyl (optionally substituted by one, two or three substituentsindependently selected from halogen, hydroxyl, and C₁₋₄alkoxy).
 17. Thecompound of claim 9, wherein R₂ is selected from the group consisting ofphenyl and a 5-6 membered heteroaryl having one or two heteroatoms eachselected from N, S, and O, wherein R₂ is optionally substituted by oneor two substituents each independently selected from the groupconsisting of halogen, and C₁-C₄ alkyl (optionally substituted by one,two or three halogens.
 18. The compound of claim 9, wherein R₂ isphenyl.
 19. The compound of claim 9, wherein R₂ is phenyl is substitutedwith one or two R₅, wherein each R₅ is independently selected from thegroup consisting of optionally substituted C₁-C₁₀alkyl, optionallysubstituted C₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl, andhalo.
 20. The compound of claim 9, wherein R₂ is selected from the groupconsisting of optionally substituted thienyl, optionally substitutedfuranyl and optionally substituted pyridinyl.
 21. The compound of claim19, wherein R₂ is a para-substituted phenyl.
 22. The compound of claim19, wherein R₂ is phenyl is substituted one or two R₅, wherein each R₅is independently selected from the group consisting of optionallysubstituted C₁-C₁₀alkyl, optionally substituted C₂-C₁₀alkenyl,optionally substituted C₂-C₁₀alkynyl, and halo.
 23. The compound ofclaim 9, wherein R_(4a) is selected from the group consisting ofoptionally substituted C₁-C₆ alkyl, optionally substitutedC₃-C₇cycloalkyl, phenyl, OR_(c), C(O)OR_(c), C(O)R_(c), optionallysubstituted heterocycle and optionally substituted heteroaryl, whereinR_(c) is selected, independently for each occurrence, from the groupconsisting of H and C₁₋₆alkyl.
 24. The compound of claim 9, whereinR_(4a) is a heterocycle, or a 5-6 membered monocyclic or a 8-10memberedbicyclic heteroaryl having one, two or three heteroatoms selected fromN, S or O, wherein the heterocycle or heteroaryl are optionallysubstituted by one, two or three substituents independently selected foreach occurrence from the group consisting of halogen, C₁₋₆alkyl(optionally substituted by one, two or three substituents eachindependently selected from halogen and hydroxyl), C₁₋₆alkoxy(optionally substituted by one, two or three halogens), hydroxyl, andNR_(d)R_(d) wherein R_(d) is independently for each occurrence selectedfrom H and C₁₋₄alkyl, or the two R_(d)s taken together with the N towhich they are attached form a heterocyclic ring.
 25. The compound ofclaim 9, wherein R_(4a) is selected from the group consisting of:

wherein each X is independently O, S or NR_(g); each R_(g) isindependently selected from the group consisting of hydrogen, C₁-C₄alkyl, C₃-C₆cycloalkyl, and each R₆, R₇and R₈is independently selectedfor each occurrence from the group consisting of hydrogen, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₁₆ alkynyl, C₃-C₇cycloalkyl, C₃-C₇cycloalkenyl,phenyl, heterocycle, heteroaryl, halo, hydroxyl, carboxyl, OR_(c),NR_(d)R_(d), C(O)OR_(c), CN, C(O)R_(c), wherein the C₁₋₆alkyl, C₂-C₆alkenyl, C₂-C₁₆ alkynyl, C₃-C₇cycloalkyl, C₃-C₇cycloalkenyl, phenyl,heterocycle, and heteroaryl of R₆, R₇and R₈may each be optionallysubstituted by one, two or three substituents selected from halo,hydroxyl, C₁₋₆alkyl and C₁₋₆alkoxy; R_(c) is C₁₋₄alkyl; and R_(d) isindependently for each occurrence selected from the group consisting ofH and C₁₋₄alkyl, or the two R_(d)s taken together with the N to whichthey are attached form a heterocyclic ring.
 26. The compound of claim23, wherein R_(4a) is an optionally substituted C₃-C₇cycloalkyl.
 27. Thecompound of claim 26, wherein R_(4a) is an optionally substitutedcyclopropyl or an optionally substituted cyclobutyl.
 28. The compound ofclaim 26, wherein R_(4a) is a C₃-C₇cycloalkyl substituted with asubstituent having the formula:

wherein: each R_(h) is independently selected from the group consistingof hydrogen, halo, optionally substituted C₁-C₁₀alkyl, and optionallysubstituted C₃-C₆cycloalkyl, or two geminal R_(h) groups areindependently taken together with the carbon atom to which they areattached to form an optionally substituted heterocyclic or an optionallysubstituted heteroaryl; R₉ is selected from the group consisting ofhydrogen, optionally substituted C₁-C₁₀alkyl, optionally substitutedC₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl, optionallysubstituted C₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl,optionally substituted aryl, halo, OR_(c), NR_(d)R_(d), C(O)OR_(c), NO₂,CN, C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d), NR_(d)C(O)R_(c),NR_(d)S(O)_(n)R_(c), NR_(d)(COOR_(c)), NR_(d)C(O)C(O)R_(c),NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)NR_(d)R_(d), NR_(d)S(O)_(n)R_(c),S(O)_(n)R_(c), S(O)_(n)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c),optionally substituted heterocyclic and optionally substitutedheteroaryl; p is 0, 1, or 2; and wherein the C₃-C₇cycloalkyl isoptionally further substituted.
 29. The compound of claim 23, whereinR_(4a) is selected from the group consisting of:

wherein each R₁₀is independently selected from the group consisting ofhydrogen, optionally substituted C₁-C₁₀alkyl, optionally substitutedC₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl, optionallysubstituted C₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl,optionally substituted aryl, halo, OR_(c), NR_(d)R_(d), C(O)OR_(c), NO₂,CN, C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d), NR_(d)C(O)R_(c),NR_(d)S(O)_(n)R_(c), NR_(d)(COOR_(c)), NR_(d)C(O)C(O)R_(c),NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)NR_(d)R_(d), NR_(d)S(O)_(n)R_(c),S(O)_(n)R_(c), S(O)_(n)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c),optionally substituted heterocyclic and optionally substitutedheteroaryl; alternatively, two geminal R₁₀groups are taken together withthe carbon atom to which they are attached to form a spiroC₃-C₁₂cycloalkyl, a spiro C₃-C₁₂cycloalkenyl, a spiro heterocyclic, aspiro aryl or spiro heteroaryl, each optionally substituted; or yetalternatively, two vicinal R₁₀groups are taken together with the carbonatoms to which they are attached to form a fused, optionally substitutedcyclic group selected from the group consisting of C₄-C₈cycloalkyl,C₄-C₈cycloalkenyl, 4- to 8-membered heterocyclic, aryl and heteroaryl,each optionally substituted; or further alternatively, two R₁₀groupsattached to non-adjacent carbon atoms are taken together with the carbonatoms to which they are attached to form a bridged cyclic group selectedfrom the group consisting of C₄-C₈cycloalkyl, C₄-C₈cycloalkenyl, and 4-to 8-membered heterocyclic, each optionally substituted; each R_(h) isindependently selected from the group consisting of hydrogen, halo,optionally substituted C₁-C₁₀alkyl, and optionally substitutedC₃-C₆cycloalkyl, or two geminal R_(h) groups are independently takentogether with the carbon atom to which they are attached to form anoptionally substituted heterocyclic or an optionally substitutedheteroaryl; R₉ is selected from the group consisting of hydrogen,optionally substituted C₁-C₁₀alkyl, optionally substitutedC₂-C₁₀alkenyl, optionally substituted C₂-C₁₀alkynyl, optionallysubstituted C₃-C₁₂cycloalkyl, optionally substituted C₃-C₁₂cycloalkenyl,optionally substituted aryl, halo, OR_(c), NR_(d)R_(d), C(O)OR_(c), NO₂,CN, C(O)R_(c), C(O)C(O)R_(c), C(O)NR_(d)R_(d), NR_(d)C(O)R_(c),NR_(d)S(O)_(n)R_(c), NR_(d)(COOR_(c)), NR_(d)C(O)C(O)R_(c),NR_(d)C(O)NR_(d)R_(d), NR_(d)S(O)_(n)NR_(d)R_(d), NR_(d)S(O)_(n)R_(c),S(O)_(n)R_(c), S(O)_(n)NR_(d)R_(d), OC(O)OR_(c), (C═NR_(d))R_(c),optionally substituted heterocyclic and optionally substitutedheteroaryl; p is 0, 1, or
 2. 30. The compound of claim 9, wherein Y isS, S(O)₂ or S(O)₂NR_(d).
 31. The compound of claim 9, wherein R_(4b) isa heterocycle or a 5-6 membered monocyclic or a 8-10membered bicyclicheteroaryl having one, two or three heteroatoms selected from N, S or O,wherein the heterocycle or heteroaryl are optionally substituted by one,two or three substituents independently selected for each occurrencefrom the group consisting of halogen, C₁₋₆alkyl (optionally substitutedby one, two or three substituents each independently selected fromhalogen and hydroxyl), C₁₋₆alkoxy (optionally substituted by one, two orthree halogens), hydroxyl, and NR_(d)R_(d) wherein R_(d) isindependently for each occurrence selected from H and C₁₋₄alkyl, or thetwo R_(d)s taken together with the N to which they are attached form aheterocyclic ring.
 32. The compound of claim 9, wherein R_(4b) isselected from the group consisting of furanyl, pyridinyl, pyrazinyl,pyrazolyl, imidazolyl, isoxazolyl, triazolyl, thiazolyl, oxadiazolyl,thiadiazolyl, thienyl, piperazinyl, and benzimidazolyl, each optionallysubstituted.
 33. A compound selected from the group consisting of:

and pharmaceutically acceptable salts thereof.
 34. A pharmaceuticalcomposition comprising a compound of claim 1 and a pharmaceuticallyacceptable carrier or excipient.
 35. The pharmaceutical composition ofclaim 34, wherein the composition further comprises at least oneadditional CFTR modulator.
 36. The pharmaceutical composition of claim34, wherein the composition further comprises at least two additionalCFTR modulators.
 37. A method of enhancing cystic fibrosis transmembraneconductance regulator (CFTR) activity in a subject in need thereofcomprising administering to said subject an effective amount of acompound of claim 1, or a pharmaceutical composition of claim
 34. 38.The method of claim 37, wherein the activity of a mutant CFTR isenhanced.
 39. The method of claim 38, wherein the mutant CFTR isselected from the group consisting ΔF508, S549N, G542X, G551D, R117H,N1303K, W1282X, R553X, 621+1G>T, 1717−1G>A, 3849+10kbC>T, 2789+5G>A,3120+1G>A, I507del, R1162X, 1898+1G>A, 3659delC, G85E, D1152H, R560T,R347P, 2184insA, A455E, R334W, Q493X, and 2184delA CFTR.
 40. The methodof claim 39, wherein ΔF508CFTR activity is enhanced.
 41. The method ofclaim 37, wherein the subject is suffering from a disease associatedwith decreased CFTR activity.
 42. The method of claim 41, wherein thedisease is selected from the group consisting of cystic fibrosis,congenital bilateral absence of vas deferens (CBAVD), acute, recurrent,or chronic pancreatitis, disseminated bronchiectasis, asthma, allergicpulmonary aspergillosis, chronic obstructive pulmonary disease (COPD),chronic sinusitis, dry eye disease, protein C deficiency,A-β-lipoproteinemia, lysosomal storage disease, type 1 chylomicronemia,mild pulmonary disease, lipid processing deficiencies, type 1 hereditaryangioedema, coagulation-fibrinolyis, hereditary hemochromatosis,CFTR-related metabolic syndrome, chronic bronchitis, constipation,pancreatic insufficiency, hereditary emphysema, Sjogren's syndrome,familial hypercholesterolemia, I-cell disease/pseudo-Hurler,mucopolysaccharidoses, Sandhof/Tay-Sachs, Crigler-Najjar type II,polyendocrinopathy/hyperinsulemia, Diabetes mellitus, Laron dwarfism,myleoperoxidase deficiency, primary hypoparathyroidism, melanoma,glycanosis CDG type 1, congenital hyperthyroidism, osteogenesisimperfecta, hereditary hypofibrinogenemia, ACT deficiency, Diabetesinsipidus (DI), neurophyseal DI, nephrogenic DI, Charcot-Marie Toothsyndrome, Perlizaeus-Merzbacher disease, Alzheimer's disease,Parkinson's disease, amyotrophic lateral sclerosis, progressivesupranuclear palsy, Pick's disease, Huntington's disease,spinocerebellar ataxia type I, spinal and bulbar muscular atrophy,dentatorubral pallidoluysian, myotonic dystrophy, hereditaryCreutzfeldt-Jakob disease (due to prion protein processing defect),Fabry disease, and Straussler-Scheinker syndrome.
 43. The method ofclaim 42, wherein the disease is cystic fibrosis.
 44. The method ofclaim 43, wherein the subject is a human patient.
 45. The method ofclaim 44, further comprising administering an additional CFTR modulator.46. The method of claim 44, wherein at least two additional CFTRmodulators are administered.
 47. The method of claim 45, wherein atleast one CFTR modulator is a CFTR corrector or potentiator.
 48. Themethod of claim 47, wherein the CFTR corrector is selected from thegroup consisting of VX-809, VX-661, VX-152, VX-440, GLPG 2665, GLPG-2222and VX-983 and the CFTR potentiator is selected from the groupconsisting of GLPG-1837, ivacaftor and genistein.
 49. The method ofclaim 48, wherein one of the at least two additional therapeutic agentsis a CFTR corrector and the other is a CFTR potentiator.
 50. A method ofidentifying a candidate agent that increases CFTR activity, comprising:a) contacting a cell that expresses a CFTR protein with the candidateagent and a compound of claim 1; b) measuring the CFTR activity in thecell in the presence of the candidate agent and the compound of claim 1;and c) comparing the CFTR activity to that in the absence of the testagent, wherein an increase in CFTR activity in the presence of the testagent indicates that the agent increases CFTR activity.
 51. The methodof claim 50, wherein the cell expresses a mutant CFTR protein.
 52. Themethod of claim 50, wherein CFTR activity is measured by measuringchloride channel activity of the CFTR, and/or other ion transportactivity.
 53. The method of claim 52, wherein the method ishigh-throughput.
 54. The method of claim 50, wherein the candidate agentis a CFTR corrector or a CFTR potentiator.